NO COORDINATION BETWEEN M P GOVERNMENT AND VETERINARY UNIVERSITY

Above is the head line of Today's (22nd January,2015) Dainik Bhaskar News Paper (Jabalpur) while informing about resignation of the newly appointed (joined 19th May,2014) vice chancellor Dr Amarjeet Singh Nanda who hails from Ludhiana. This Eminent scientist and  former Animal Husbandry Commissioner, Government of India is selected as Vice Chancellor of Veterinary University, Ludhiana and now preferring to leave Jabalpur. Obviously, he appeared less interested to remain at Jabalpur otherwise why he applied for vice chancellorship of Ludhiana  .While planning to leave Madhya Pradesh , he could not resist to express his feelings, though in sophisticated manner,about no coordination between Madhya Pradesh Government and  the University. This expression speaks a lot why no veterinary or agriculture university of the state is among top institutes of the country ? As JNKVV is the oldest Agriculture University of MP while Gwalior Agriculture University and Veterinary University are new entrants , we can understand the facts by knowing 50 years history better than commenting on the new universities whose outcomes are still not well known ; more over these new universities had been old constituent  colleges ,once upon a time, of JNKVV   ( e.g. Jabalpur veterinary college was part of JNKVV except few years back ) .

PANTNAGAR VS JNKVV :  If we go back into the history of Agriculture education in the state, the Jawaharlal Nehru Krishi Vishwa Vidyalaya, (JNKVV) Jabalpur was inaugurated on 2nd October , 1963 by Smt Indira Gandhi, the then information minister of Government of India and was  third /or second (?) on American  land grand pattern Agriculture University in the country- the first was Pantnagar University and second(?) was PAU,Ludhiana.(But JNKVV came in operation w.e.f Oct 1964 with start of  semester system for 1964-65 BVSc batch) 

Pantnagar Agriculture University was established on a single campus basis which started its own Agriculture, Veterinary colleges but did not include either old Mathura Veterinary College or Kanpur, Bichpuri, Naini Agriculture colleges into its fold .This helped in consolidating  its resources and concentrating on specified goals by encouraging newly appointed faculty ( very young PG holders became Assistant Professors- a rare opportunity at that time).
On the other hand, JNKVV included all the state agriculture colleges ( Jabalpur,Indore,Gwalior,Raipur,Rewa,Sehore) and veterinary colleges (Jabalpur, Mhow) in its fold and thereby boosted itself the largest Asian Agriculture University.But this system worked as if JNKVV is an affiliated university and  inadvertently became responsible  for deteriorating education standards in other campuses . This was mainly because earlier all the Agriculture and Veterinary Colleges were having their own state level Professors in each department (generally three faculty in each dept- 1 professor, 1-2 Asstt Professor, 2-3 demonstrators) but with the formation of JNKVV they were reduced to Associate Professor level (many demonstrators, assistant professors preferred to go back to Veterinary department). Perhaps, greater harm was done by creating only one post of University Professor of the subject and that was allotted to Jabalpur , being main campus of the university, bringing all earlier state level professors under them - obviously they considered it as demotion and this could not auger well education and research atmosphere in outside campuses whose head was located  at Jabalpur. Further JNKVV was not allotted matching finances that can take up salary burden of enlarged staff as well as could create infrastructure for research and advance education.  

APPOINTMENT OF EMINENT PROFESSORS : It is not that attempts were not made to attract talented faculty members (from outside of Madhya Pradesh) to join as University Professors at Jabalpur. I still remember that Dr A Ahmed joined professor of Pharmacology, Dr PN Bhat, Genetics, Dr Ranjan , Nutrition, Dr BS Malik , Microbiology, Dr SC Dutt, Parasitology etc . Ironically, all (except Dr BS Malik) the professors left veterinary college Jabalpur one by one and as the history tells all of them attained important positions in veterinary profession confirming their inherited talent that could not be explored in M.P or  JNKVV . If you ask the reason of leaving Jabalpur , privately they will tell Jabalpur is a dead place where there is no atmosphere (or infrastructure) for research or expression of talents. These eminent professors were not vocal in voicing shortcomings (or more wise ?)  and considered it befitting to leave  JNKVV and join elsewhere. The best example I may cite is of Dr SC Dutt,(IVRI scientist)   who was a Rafi Ahmed Kidwai award winner , my Guru ( he guided me in my MVSc program ) and was really interested in teaching and research. But he also left JNKVV within four years and joined as ADG (education) at ICAR but did not like that job and was taken as Professor and Head of newly formed department of Parasitology, Ludhiana by Dr BS Gill, the then Dean faculty of PAU. I do not think if there have been any serious thinking why eminent teachers do not remain in JNKVV or the administration has ever  tried to improve  atmosphere of JNKVV .

JNKVV VICE CHANCELLORS : Its true that the state government tried to bring eminent persons of the country to join as vice chancellor of the university- first was Dr HM Patel, then Dr Negi , Dr SukhDev Singh. Sadly, none of them completed full term of vice chancellorship and left the university prior completing the term. Dr Chandraka Thakur from Bihar had controversial tenure of vice chancellorship in JNKVV and has to leave Jabalpur prior completing his term. As no outside vice chancellor remained in the university for full term or have shown improvement in teaching and research , the teachers of JNKVV agitated to appoint a local teacher as its vice chancellor with the slogan we want ' son of the soil'. To this one professor , sarcastically commented  that we should ask for son of the soil but not  night soil.

FREEDOM OF WORK OR AUTOCRACY :  As teaching and research is a highly creative profession, it can flourish only where a teacher considers that his genuine grievances are addressed correctly by a right authority and that he is not victimized and his talents are recognized by the authorities  . Unfortunately, this is the most important fact which JNKVV has failed to impress to  its faculty . Here the Board of Management, the highest executive authority , did not have any faculty as its member hence there was no method to verify vice chancellor's provisions . Secondly, Board's proceedings are confidential and no employee of JNKVV can meet its board member to raise its grievances as this will be treated against service conditions. Thus if you are aggrieved with university decision (or vice chancellor in real term) you have to apply to the Chancellor of JNKVV who remains mute in most of the  cases hence ultimately the employee takes  High court's shelter.This is the reason why there is maximum number of court cases in the university. In such circumstances , the vice chancellor becomes an autocrat , some time a whimsy administrator whose only aim becomes to pass smoothly his tenure .
No doubt vice chancellors should be given freedom to work without any interference from state government but simultaneously there should be a check so  that vice chancellor should behave in a transparent way . Certainly, JNKVV is not their private company. Unfortunately actions of the vice chancellor have never been transparent or beyond personal prejudice  .  I am referring only a few examples from the past which clearly show  functioning of JNKVV :

• When a vice chancellor was disturbed  by the President of Technical (teachers) Staff Association  he terminated him with out any such power . The vice chancellor is alleged to claim 'ham ne to terminate kar diya, ab ho ne do paresan ; jyada se jyada phir se restate he to hoga".
• In a Professor's interview, the vice chancellor (who happens chairman of selection committee) was pleading to select an inferior candidate over a meritorious,national award winner candidate by stating " We have to run the university where these meritorious teachers are useless - they are of no use except teaching and research" And much inferior person was selected as Professor over a meritorious award winner person.
•  The vice chancellor does not care to follow MOU signed by himself and there is no body to look after such facts. 
• Even  the university has not hesitated to present contradictory documents in the High court thereby eroding its image before the employees i.e. when a university can file a false document in the court of law how one can believe it will always tell truth in the board of management .
• When JNKVV faced financial crunch , it circulated a circular terming  "attending seminars, conferences, symposiums " as unproductive works.

TECHNICAL WORK : This is most important aspect where no coordination between state government and the university is visible. Its true that prior establishing JNKVV, all the veterinary and agriculture colleges were under their respective government departments. The main or only function of these colleges was teaching the subjects and there were frequent transfers of the staff between the department and colleges. But with the formation of JNKVV, there became a water tight compartment between  the department and college/universities. This was mainly because of requirement of master's or doctorate degree for university posts while these were not essential for departmental posts or promotions. 

However, this water tight compartment also resulted in no transfer of technology ,developed in the university to the department. It appeared that the department has no concern with what is developed in the university , neither teachers took interest in what is going on in the field. Rather , a miss trust developed between the two. While the field staff discarded all the scientific developments in JNKVV as 'academic exercise' not applicable in the field, the university staff alleged that field staff is not following technical knowledge while dealing the subject. And this no coordination between MP Government and Veterinary University is visible by many of those who have concerns with development of the state. I do not remember if  Director of Veterinary services came to the veterinary college and urged to solve some field problems or came to the staff to transfer some technology to the field. It may be pertinent to provide some examples where scientists developed the techniques but no government official ever cared to arrange for its transfer in the field or to improve the same :

Dr ML Mehta, Professor of Medicine and his associates developed a crude vaccine to protect the poultry against spirochaetosis ,  but this was not transferred in the field.
Dr BS Malik  ,Professor of Microbiology and his associates isolated a strain of Rani Khet virus which was important in protecting poultry against the disease (for this, they were awarded Hari Om Ashram Award,ICAR) but state vaccine production center did not come forward for its field application .

Dr AG Khan, Professor of Poultry Science developed "Narmada XL" a colored bird for backyard poultry (again received ICAR awrad for this research work) but this was not taken up by the department to popularize in the villages.

I was awarded ICAR National Fellowship to work on different aspects of schistosomiasis. This work resulted in publication of a book on "Schistosomes and schistosomiasis in South Asia" with development of many new techniques.This work proved "Hatching method" a simple, cost effective method of diagnosis and triclabendazole a better proposition for treating the infection, in place of Anthiomaline. But till date , no one from the government side tried to contact me for transferring the knowledge to the field.

OTHER STATES : This situation does not appear to exist in all the states. There are states where a better coordination is existing and the government is fully utilizing the services of university staff. The chief minister of Haryana often called the staff of HAU to solve one or other problem of the state. Generally, one may notice one or two ministers visiting frequently HAU for consulting in technical matters. So is the case with Punjab or TamilNadu.

Even Rajasthan Government is telling their requirements and asking the staff to carry out research work in particular field. These are the efforts of the university staff and veterinary directorate that a book on "Anatomy of Camel" and other books on animal husbandry were published for the benefit of animal owners. 

Perhaps, Dr Nanda might be having  above feelings while expressing no coordination between MP Government and Veterinary University. One fact is clear from this expression. The state  can get only  a little  by appointing an eminent scientist as Vice Chancellor of a Veterinary/Agriculture University, unless until it does not review its action plan and does not improve coordination between state government and the university , as is happening in some progressive states. 

STRATGEY TO CONTROL PARASITIC INFECTIONS IN CHHATTISGARH STATE

 This is the write up discussed in the  BRAIN STORMING WORKSHOP held on 12th May 2014 at Chhattisgarh Kamdhenu Vishwa Vidyalaya,Durg and appeared in the Souvenir on page 49-51. These are the practical suggestions that may certainly reduce parasitic burden of a locality hence any one may try in other places too but of course giving due credit for the idea as honesty desires . It will be my pleasure to help any one in this endeavor.

MC AGRAWAL

Board Member, Chhattisgarh Kamdhenu Vishwa Vidyalaya, Durg  

It is my great privilege and honor  that I have witnessed great eagerness in our Honorable Governor Shri Shekhar Dutt jee as  how he can take  Chhattisgarh to new heights- how economy and health of its people can be enhanced tremendously . Actually, this brain storming workshop is his brain child, whose deliberations will pave the way to map out new strategies for development of the state. I consider myself privileged and lucky to  be a part of this whole program . Looking to my experiences and also importance of the subject to the state , I have selected above topic. Here, I will not discuss academic matter but only those issues which have direct bearing in controlling parasitic  infections in the state.

1.       ANY  GOVERNMENT PROGRAM ON CONTROL OF  ANIMAL PARASITES; The central government is running various disease control programs in humans like control of malaria, tuberculosis, polio , lymphatic filariasis. Even in animal’s health, there is control program on Rinder-pest, foot and mouth disease etc where a lot of funds, technical help, men power are allotted .

Is there any comparative program of central government to control any parasitic disease of any animal species ? As per my knowledge there is none.  Why it is so ?

2.        LITTLE OR NO HARM FROM PARASITES ? Possibly, the parasitic infections in animals have been neglected being considered causing little or no harm to the animals. This notion came ,perhaps, by comparing the mortality rate caused by bacterial ,viral infections and those by parasitic infections. No doubt,  there is difference in mortality rate between these infections but many times  our diagnosis has  been faulty and  failed to identify parasitic etiology of death as has happened in two outbreaks  in sheep in Karnataka and Andhra Pradesh states (Chandra et al 2003 Indian J Vet Pathol 27:93-94);  In one outbreak, 80 sheep out of 400 died while 50 others showed  symptoms including swollen temporal region ,eye walls and diarrhea with mucus and blood. In one outbreak the disease was suspected  PPR ( Peste de petits ruminants) while in other Johne’s disease- but  parasitic disease in none . However, diagnosis ,carried out by IVRI centre for animal disease  research and diagnosis, revealed the death ,in both outbreaks,  due to  ovine schistosomiasis (Schistosoma indicum). How frequently this faulty diagnosis is occurring under field conditions ? we do not know ? Does it not reflect our pre-conceived notions regarding parasitic infections and also how faulty is our diagnosis at field level ?

3.       DIAGNOSIS OF PARASITIC INFECTIONS IN FIELD LABORATORIES :  The above example showed that field laboratory could not identify schistosome eggs in the fecal material .This is since our field veterinarians are  posted not as per their specialization but casually and this is also happening in our veterinary diagnostic laboratories. It is a fact that our veterinarians are more conversant about diagnosis of bacterial and viral infections but have little expertise in diagnosing parasitic diseases. I had an occasion to check registers of some diagnostic laboratories and surprised to note their results ; it was  like – positive for helminth eggs, positive for nematode/fluke eggs ,for coccidia ;they may identify trypanosomes in a blood slide but difficult for many to differentiate Babesia or Theileria organism from stain granules ; they can identify coccidian oocysts but could not its merozoites –the stage where damage is being carried out. We are not sure how many are trained about new techniques related to diagnosis of pathogens. And then, the expert diagnostician is transferred to some other place -to be replaced by a poultry  or alike expert.

Obviously, this diagnosis  pattern has to be changed ,if we plan to know the harm caused by the parasites . This needs  recognition of specializations while posting veterinarians in diagnostic laboratories, their training and overall monitoring of the results proclaimed by the diagnosticians .

4.       LACK OF HOSPITAL DATA: If we want to assess correctly damage caused by any ailment , our veterinary hospitals should record each animal case in such a way that the same may be assessed and analyzed  by any researcher  . Presently, the veterinary hospitals are recording the cases in a register which is difficult to analyze.

5.       ECONOMIC ASSESSMENT OF DISEASE LOSSES : This leads us to a pertinent question how we assess economic losses due to animal diseases. This loss should not be confined only to mortality but also due to other losses like milk production, body weight loss, wool quality ,reproduction etc  and then only we will be able to know the losses made by each disease in each host species. I am glad to inform this gathering that Chhattisgarh Kamdhenu University has resolved to start a separate department of veterinary economics and statistics .This department  will assess economic losses occurring in animal industry in the state and will be able to generate data  related to animal industries.

6.       HOW WE MAY PLAN CONTROL PROGRAM : Keeping above facts in mind and also that we do not have any commercial vaccine for any parasite or a large number of drugs against any parasite , we have to plan both short and long terms for controlling  parasitic infections  in Chhattisgarh state.

7.       STEREOSCOPIC MICROSCOPE IN DIAGNOSTIC LABORATORIES : Those who are familiar with parasitic infections are well aware about the importance of a stereoscopic microscope in identifying free living parasitic stages like larvae, cercariae, snails, ticks, insects  and other parasitic forms which are essential in parasitic diagnosis. But our field veterinary laboratories are  lacking a stereoscopic microscope thus impeding parasitic diagnosis . The least we can do is to provide a good stereoscopic microscope in every diagnostic laboratory.

8.       EXPERT DIAGNOSTICIAN : Now time has come when we should start recognizing importance of specializations in veterinary field . The veterinarians should be posted as per their post-graduate qualifications. Thus , diagnostic laboratories should be staffed only with those holding post graduate degrees in Pathology, Parasitology or Microbiology. They should be well aware about diagnosis of important parasitic infections or may be trained for the same.

9.       MANAGEMENT IN ANIMAL HUSBANDRY : In the present scenario, management in animal husbandry is most important in controlling parasitic infections and following points may be tried with the condition of analyzing effect of each factor so that their importance may be identified :

10.   GOBAR GAS PLANT OR VERMICOMPOSE : If we start collecting cow-dung from all the places and use it either in gobar gas plants or in preparing vermicompose, this single step will not only improve economy of rural household but  curtail parasitic infections to a great extend since almost 90% parasites excrete their infective stages through feces (we have to devise alternative methods for other animal species ).

11.   NO OVER USE OF MEDICINES AGAINST PARASITIC INFECTIONS :  Presently chemotherapy is the only remedy used by our field veterinarians against parasitic infections and to a greater extend there is indiscriminate use of the medicines. We do not have large number of drugs against parasitic infections ; neither research is promoted for developing new drugs. There are reports of developing drug resistance in the parasites and if it increases further, we will not have effective drugs against dreaded parasites. Therefore, there is the need to use the drugs judiciously while dealing parasitic infections as well as monitoring drug resistance in the parasites.

12.   HOW TO USE FAMACHA SYSTEM OF FAO : The organization has developed a method of assessing anemia in small ruminants by comparing paleness of conjunctiva  and recommending treatment, against Haemonchus Contortus infection, of the animals showing anemia, without further examination . However, this system is developed for the countries where only Haemonchus contortus is a problem. Since our country has a multiple parasitic problems, each having different treatment, this system can not be followed in Toto. Instead of taking anemia as the only criterion, I suggest to check animals for anemia, diarrhea , bottle jaw  ; if any of these symptoms are present in the animal, their excreta should be examined by laboratory  methods for parasitic infections and positive cases should be treated accordingly.

13.   TRIBAL EXPERIENCES AND SEARCH OF  MEDICINAL PLANTS : There is a need to study how tribes are treating their animals against important ailments. As tribal population in the state is more than fifty percent, their rich experience should be exploited  which may lead us identifying some new plants having rich anti-parasitic activities.

14.   COLLABORATION WITH OTHERS : There are many stakeholders who are working on animal diseases either directly or indirectly but are working in isolation. The university is not an executive body hence it is important to have a close link with executive body i.e. animal husbandry department. Likewise, there is a need of collaboration with public health institutes particularly for preparing strategies for zoonotic diseases and all the stakeholders must work in collaboration for controlling communicable diseases. I have already emphasized the need for a National Institute on Parasitology (www.indianschistosomiasis.blogspot.com) .

15.   USE OF INTERNET /VIDEOS /MOBILES : We are living in twenty first century which is witnessing spread of internet and mobiles with high speed. Therefore, it will be prudent on the part of the university/AH Department to  prepare short videos of about 5-10 minutes depicting important control strategies , diagnosis methodologies with photographs of the parasitic stages and to up-load the same on different websites including you tube. These devises should also be used to redress any problem ,faced by field staff or animal owners.

16.   PRIORITIES IN OUR RESEARCH AREAS  :  We have to reschedule priorities of our research areas which should help in controlling parasitic infections. All the above fields require constant monitoring so that best results may be identified. Thus , there is the need to assess importance of each animal species (with its age and sex) in spreading one or other parasitic infection in each geographical area ; to check efficacy of laboratory diagnostic techniques , ante-mortem and post mortem, in diagnosing parasitic infections; developing new parasitic techniques for diagnosis( but  can not over-emphasize immuno-diagnosis) ; economic losses at every stage of the infection, knowing local medicinal plants for curing parasitic infections and other new research fields . Here, I will like to mention following points also ,regarding our research priorities :

(a).CLINCAL NUTRITION : This aspect has been neglected in veterinary medicine and a new course on “Clinical Nutrition” should be introduced at post-graduate level . This will have two parts. The first part is about the nutrition like protein diet ,minerals, vitamins etc which help in increasing resistance against the disease while second part, less studied , is about the deficiencies caused by the diseases at different times and the need of supplementing these constituents while treating the animal for particular disease.

(b). PARASITIC SUSCEPTIBILITIES : Generally, the animals are found infected , in nature, not with one parasitic infection but a multiple parasitic infections. Many times, it has been difficult to pin point pathogenic effect due to  a specific  parasitic species.

Again, there are reports that infection of one parasitic infection makes animal susceptible to another important parasitic species. For example, in one experimental study at IVRI sheep , infected with Schistosoma indicum, were found harboring three times more  number of Haemonchus contortus  in its abomasum  in comparison to the control sheep and the difference was attributed to schistosome infection  (Srivastava et al ,1964 Indian J Vet Sci 34:35-40). Obviously, such studies are very few but require our attention.

(c ) MENTION OF PARASITIC SPECIES NOT EXISTING IN INDIA : In the technical or semi technical writings , I have witnessed mentioning of many parasitic species, by the veterinarians , that are not existing in India and which are even difficult to establish in India due to absence of particular intermediate host (at times, it creates problems to other stakeholders ). Thus , I have seen the papers about existing of Schistosoma japonicum or Fasciola hepatica (in non- hilly region ) , Trypanosoma bruci or alike ones which are not existing in the country. A some what similar problem is  in mentioning drugs/ molluscicides /insecticides  that are absent from  Indian markets.

If we scrutinize the problem ,it appears that at undergraduate level , the teachers fail to emphasize about the parasitic species which are not existing in the country. As our VCI has reduced undergraduate courses in parasitology, will it not be a good proposition to teach , at undergraduate level, only those parasites which are existing in the country ? And in post-graduate courses details of all parasites should be taught so that scholars should understand epizootiology of each infection, very clearly.     

In my opinion, professor level scientists should be given hard research areas and we must discard the research which has continuously proved  unfruitful .

WITH PRESENT KNOWLEDGE , CAN WE SUCCEED IN CONTROLLING SCHISTOSOMIASIS IN INDIA

This is the second lecture delivered in the winter course at IVRI,Izatnagar on 11th Dec 2013 and i am reproducing the same here . I will appreciate to receive any comments which may further the subject .It will be my pleasure if any research organisation is interested to collaborate in this endeavor. 

                                                                M C AGRAWAL

                              FORMER NATIONATIONAL FELLOW AND EMERITUS SCIENTIST

                                 COLLEGE OF VETERINARY SCIENCE AND A H , JABALPUR 482001

                                                             drmcagrawal@gmail.com    

It is heartening to note that one part of this winter course is to teach molecular techniques about control of parasitic diseases .In the past also, there had been some advance courses and discussion about control of parasitic diseases . But whether these discussions and advance courses can lead to  control of parasitic diseases ? Obviously, the answer is no. Now, with the changing time, it becomes imperative to know why we need to control parasitic diseases, which diseases are given priorities , and how we can control  these parasitic diseases.  And this is what I am going to discuss ; thus my lecture will not be a mere academic exercise of citing references  from some research papers. There is now time ,we should discuss how we may try to control parasitic diseases and what should be our research priorities that may help in achieving these goals.   

                If we analyze disease control program of our country, it will be clear that the country has given top priority ,logically ,for controlling those infectious diseases that are affecting human population. Thus there are central government’s programs for controlling tuberculosis, malaria, filariasis, guinea worm or naru  ,polio (small pox already eradicated ) etc Interestingly, almost no attention is paid on zoonotic aspects of these diseases  or medical faculty has  not taken diseases where animals are playing a crucial role in their maintenance. For example, we have not taken sincere  steps for controlling rabies .This reflects how wide apart are the two professions (medical and veterinary) which are supplementary to each other  to combat zoonotic infections of the country.

                When we come to animal diseases ,the control programs have been launched only to limited diseases like Rinder-pest, FMD, anthrax with vaccination programs of these and other infectious animal diseases. These are also the diseases ,where the veterinary hospitals ,districts and states generate the data and pass on to government of India.

                Has government of India ever launched any control program related to any parasitic disease of our domestic animal ? Here I mean control program like that of Rinder-pest or FMD . To my knowledge, there was or is  none. No doubt there have been some vaccination programs for controlling parasitic diseases . The one was about lung-worm vaccination for controlling lung-worm infection of sheep and goats ,particularly in Jammu & Kashmir , Himachal Pradesh ( now, vaccine production has been stopped by IVRI though infection is still existing ).  The other is Theileria vaccine produced by NDDB and supplied for controlling theileriosis among cross bred cattle . Again , this is not a infection which is affecting all our domestic animals. Thus we have attempted to control a infection (lung-worm) which is restricted to certain states or a infection which is affecting limited number of our domestic animals. And we have shied  away in paying our attention to the parasitic diseases which are affecting almost all domestic animals and existing in  all the states. Therefore, we have come to a crucial point where we need answer of above questions - why we need to control parasitic diseases, which diseases are given priorities , and how we can control  these parasitic diseases.  And my this short lecture will try to discuss some of these points with the hope that you , our young Parasitologists ,will devote some time to find answers of these and related  questions which are important for controlling parasitic diseases in the country. Answer of these questions are important as you will agree with me that all your research efforts will not fructify unless and until you attempt to control the parasitic infections in our country, thereby reduce mortality and animal production losses.

WHERE IS THE DATA :

If you go to any funding agency or even any government, the obvious question will be why we wish to control the particular parasitic disease. Naturally, you will put up the data about mortality and pathology of that parasitic disease . By the way, from where have you collected these data ? From some research paper or a review article or from a book ?  And how they have calculated these data  ?

I am sure that your data have not been collected from veterinary hospitals of the state or Animal Husbandry department of the country . And when these data have not been collected from the villages whether they are representing correct scenario of the infection is debatable.

VETERINARY RECORDS :

What I have said above, is all because of my experiences . While working on my book “Schistosomes and schistosomiasis in South Asia” (Agrawal,2012) ,I tried to collect the data of nasal and hepatic schistosomiasis from veterinary hospitals and veterinary diagnostic laboratories (division level),state and central governments ;  as ,in my opinion, compilation of data from these sources is important since research journals fail to provide a detailed view of the infection ; these journals have not reported each and every outbreak or prevalence of the infection in each animal species from each geographical area .

                To my dismay, I could not collect any data, related to schistosomiasis, from any of above  source ,details have been discussed in my book .Here, it will be suffice to mention :

·         There is no direction from the government to keep record of parasitic diseases and details thereof. Neither any data are passed to state or central government .

·         The veterinary hospital registers are maintained in such a way that you cannot extract any information from them. Even it is difficult to differentiate between new and repeat cases, cattle and poultry  (I have written about this topic which has been published in the news letter of National Academy of Veterinary Sciences when Dr S K Ranjan was the President).

·         The veterinary diagnostic laboratories are vague in their reporting and counter checking is missing . While reporting coprological results (generally by direct wet smear method ) ,generally positive cases are reported  as “positive for helminth or fluke or roundworm eggs” . Thus they have not differentiated the eggs where it was easy to differentiate .

It is clear that we need changes in above system and a fresh outlook  is needed to the problem so that  we will be able to  collect the data related to parasitic diseases.

WHICH PARASITIC DISEASES TO CONTROL ON PRIORITY :

This is an important question  that if we decide to control parasitic diseases what should be our priorities ?

Answer of this  question , at first hand, appears simple i.e.  the parasitic disease which is causing greatest mortality and highest pathogenecity in domestic animals should be  attended first. But how we will reach to this conclusion when we have not generated data in this regards. And when we are attempting to generate the data, are we using sensitive and specific methodologies  ? This aspect is important else our results will not be accurate. While deciding importance of the disease, we have to examine its prevalence in each animal species, age and sex wise ; the seasons when disease is occurring; animal husbandry practices  and  geographical areas where infection is existing.

With the above, two more important aspects are also linked . One is the mortality occurring in the infected animals. It may happen, looking to Indian conditions, that the animal is not infected with single parasitic infection but multiple infections and in such cases we have to device methods for ascribing death etiology . The other aspect is related to pathology of the infection which is reflected to animal production losses. No doubt we will need new methodologies to decide the above facts and these may change as per advancement of our knowledge and environment. If we will not follow new techniques , there are all chances of making mistakes as has been done while reporting schistosomiasis in post-mortem cases ( Rathore,1998).

PRIORITY AMONG HELMINTHIC DISEASE :

As my present discussion is related to schistosomiasis, I may raise a question to you – which is the most important helminthic disease of our domestic animals ? Some of you will answer - haemonchosis, or gastro-intestinal nematodiasis, others may answer fasciolosis , or amphistomosis ; while some of you will say nasal schistosomiasis but  a few or none will ascribe hepatic schistosomiasis as the most important helminthic problem of our domestic animals . May I ask the reasons for your such conclusion ?

Those who are ascribing fasciolosis as most important helminthic disease will put forward their slaughter house experience where most of the livers yield adult Fasciola gigantica  .Further, the faecal examination shows presence of Fasciola eggs in many cases ( its different that we are not differentiating eggs of Fasciola from those of amphistomes ). And those who are new to Indian  parasitology will cite the reference of a foreign veterinary book where fasciolosis is considered as most important helminthic disease of domestic animals.

Those who are ascribing amphistomosis as most important will support their argument that every rumen in slaughter house possess one or other species of amphistomes and almost all faecal samples are positive for amphistome eggs.

Against these arguments ,we do not have such visibility of hepatic schistosomiasis hence can not be considered as most important helminthic disease in India. I have discussed this topic elsewhere ( Agrawal,1999 2003) but will like to draw your attention to the need of employing correct techniques for reaching a correct decision. As time does not permit me to go into details of such discussion, I will mention only silent points emphasizing how important it is to use a correct technique and unbiased attitude for reaching to a correct conclusion.  I am mentioning some facts for your consideration and further investigations :

·         Fasciola gigantica, the only existing Fasciola  species in India , spreads through Lymnaea auricularia or Lymnaea acuminata which is a strict fresh water aquatic snail (unlike to amphibian  Lymnaea truncatula –intermediate host for F.hepatica) and survives only in perennial water sources hence animals will get infection only from perennial water sources.

·         F.hepatica  is more damaging to domestic animals than F.gigantica .

·         Against one species of Fasciola, there are at-least five schistosome species (Schistosoma indicum,S.spindale,S.incognitum, Orientobilharzia dattai and S.nasale) which are affecting our domestic animal species.

·         Schistosome infections spread through Indoplanorbis exustus and Lymnaea luteola – both species are capable to survive not only in perennial water sources but also in temporary water bodies thereby giving higher geographical spread.

·         A large number of amphistome species (40-42) are existing in the country (Dutt,1980) some of which have been synonymized. However, only four genera have been incriminated in pathogenesis to domestic animals .

·         In amphistomosis , it is only young animals which suffer and that too only from immature amphistomosis.

·         Many fresh water snails are acting as intermediate hosts for amphistomes hence their geographical spread is as large as that of schistosomes.

Now you have  to determine ,using new methodologies, which helminthic disease is more important in our domestic animals. Here, you might have noted one more fact  ; this importance will vary as per our husbandry practices. All these fluke infections are important only in grazing animals with limited effect in stall fed. Thus our priorities will vary as per animal species, husbandry practices and geographical areas. Therefore, you will identify  those parasitic diseases to be given national importance and those with regional importance etc. 

HOW TO CONTROL SCHISTOSOMIASIS :

We have come to the last part of our lecture .And that is if we decide to control any parasitic disease in our domestic animal , how we can control  it . I will take references from schistosomiasis which have been discussed in more details in my book (Agrawal,2012). Indeed, this is the only helminthic disease that has been given so much importance and where so many countries agreed to launch control program in collaboration with WHO (1985) . Ironically , India has not participated in such program.  Interestingly, WHO have not touched zoonotic aspects of the infection while attempting control program of human schistosomiasis . The review of this  program may high light our short comings  which are important to get rid of for achieving any success in controlling any parasitic disease and also  animal schistosomiasis in India or South Asia.

MULTIPLE APPROACH :

No disease can be controlled by following a single step approach , how so ever,  effective it may be . The infection has to be tackled from all aspects so that prevalence of it may be reduced to the levels where further transmission becomes difficult .Following multiple approaches were followed in schistosomiasis :

·         Chemotherapy

·         Health education

·         Snail control through use of molluscicides

·         Environmental modifications

You will realize that such program is not possible without a large team and funds  which a department of Parasitology of an institute cannot afford .  No doubt , this control program of schistosomiasis was able to reduce prevalence rate as well as pathogenesis ,as the latter is directly proportion to the intensity of the infection; and the success was  both due to  political will and scientific team work . Here, it will be prudent to discuss scientific reasons for their success and how it will fit in our program of controlling animal schistosomiasis in India. One important difference has to be remembered that while following any control program ,we have to monitor  not one but more than one animal species.

CHEMOTHERAPY :

The most important reason of the success ,perhaps, was mass treatment of the population with a most effective schistosomicide (Praziquantel) whose single oral dose @ 40-60 mg/kg body weight was 80-95% effective in urinary and hepatic schistosomiasis . Since the weight of children varied between 20-50 kg, the dose of Praziquantel was cost effective, beside easy in administration .

When we compare this scenario with that of India,  our schistosome species  are different from those dealt by WHO hence there are all differences in schistosome bionomics . And praziquantel against any of these Indian schistosome species (S.indicum,S.incognitum, S.spindale ) has not proved as effective as reported against S.haematobium or S.mansoni . In fact, our experimental work on pig- S.incognitum model has failed to reduce schistosomes to a significant level  with presence of left over schistosome population as high as 40% ( Shames et al 2000)  A same scenario was observed when the drug was tried in other experimental models ,with schistosome reduction ranging from 50-65% (left over fluke population between 50-35% ) ,  but using other Indian schistosome species. Ironically, no available flukicide (including anthiomaline ) was found to lead to a significant reduction in schistosome population in any experimental model, tried in our laboratory or by other workers ( Agrawal ,2012).

Therefore, it is not advisable to use Praziquantel or any other flukicide in any mass treatment program which has failed to reduce schistosome population to a significant level in our animals. This is because such use will lead to the high probability of development of resistance of left over schistosome population against praziquantel .

One more hindrance in using praziquantel for mass treatment in animal schistosomiasis is with regards to its cost for treating bovines against schistosomiasis. In one field trial against nasal schistosomiasis in Balaghat district of Madhya Pradesh , 60 tablets of Prazi Plus^@ , which works out approximately to 20 mg/kg body wt was effective in eliminating completely clinical symptoms of snoring disease from 14 out of 18 cattle. However, cost of one treatment comes to Rs 1500/ per animal as market cost of two tablets is Rs 50/ (Agrawal, 2012) .

DIAGNOSTIC METHOD :

While implementing any control program, it is important to monitor the rate of infection and its  intensity in the given population prior and after the treatment. This monitoring is possible  by using a simple, rapid, economic but sensitive parasitological  diagnostic technique that may detect, light ,moderate and heavy infections. In WHO case , Kato-Katz technique was applied that fulfilled all the above criteria and was able even to detect light infections ( 24-96 epg) with presence of 1-4 eggs/slide, made of 41.7 mg of faeces (WHO,1985).  In contrast , a very low epg ,ranging between 2-4 in ruminants and 10-50 in pigs has been reported by almost all the Indian workers (Agrawal,2012) ; for this reason almost all egg detection methods have poor efficacy ,cannot be employed in control program, hence  warranting further work on this issue . We may also undertake further research on hatching process, as this has proved superior over egg detection methods.

 SNAIL CONTROL :

The snail control ,in WHO  program , was undertaken by using Niclosamide which has proved highly effective in killing the intermediate hosts i.e. Bulinus and Biomphalaria . Ironically , Niclosamide is not available in Indian markets and has to arrange its import from other country. We were able to procure Niclosamide and undertook  both experimental and field trials on fresh water snails , observing its high efficacy in killing Lymnaea luteola, Indoplanorbis exustus and Gyraulus but without affecting other water fauna and flora (Agrawal et al ,2005, Agrawal 2012).

However, ecological conditions of fresh water snails in India ,differ from those of other endemic countries. In India, these are the ponds, tanks and other water sources, both perennial and temporary , which are inhabited  by these fresh water snails, where as in endemic countries these were the rivers where snails are  surviving. Thus , we have more wide spread population of the snails , making our efforts more difficult while attempting snail control.

BIOLOGICAL CONTROL :

As use of chemical molluscicides may  cause environmental pollution , efforts are being made to find other alternatives for snail control . Work is being carried out to find Plant molluscicides which are more eco-friendly . Likewise, search is being made to find natural enemies of the snails that may be used as biological control agents for reducing snail population. However, it must be remembered that the biological enemy or competitor should not be introduced from outside the country/area as they may prove harmful afterwards as experienced by WHO (1985).

One natural enemy of young fresh water snails has been identified by us (Agrawal,2012) in the form of nymphets of dragonfly which co-habitat with the snails under same ecological conditions ; these nymphets are carnivorous in nature ,devouring young snails with their life span ranging from 1-3 years ( hence once introduced in the ponds, there is no need of re-introduction every year ). There is the need to develop the techniques for mass production of these nymphets which also feed on mosquito’s larvae (Agrawal 2012). Here is the importance and need of your molecular techniques which may pave the way of producing eggs and nymphets in large number , for their use in controlling snails or mosquito larvae.   

HYPER-PARASITISM  :

                As stated above,  Praziquantel  and  anthiomaline  are not able to kill a significant number of Indian schistosomes, which is not the case with S.haematobium or S.mansoni. There are also chances of development of resistance of the schistosome population  ,left over after wide use of these less effective  drugs. Therefore, there is the need to think alternatives against chemotherapy for parasitic diseases.

One promising area of research might be searching hyper-parasites of schistosomes or of  any other parasite which is causing disease in man and his animals. As you are aware these hyper-parasites are those organisms which parasitize on the parasite itself e.g. schistosomes . This hyper-parasite  may be a bacteria or a virus or even a fungus. We have to search those hyper parasites which are lethal to these parasites and should be host specific thereby eliminating our animals or human beings in causing any harm .  

NEED OF A NATIONAL INSTITUTE OF PARASITOLOGY :

With this lecture, I tried to discuss how ill equipped we are for launching a state or national level control program against any parasitic disease of our domestic animals. There is the need to re-think on our research programs and to develop methodologies which may help directly or indirectly in controlling parasitic diseases of the nation . But it will be a bad idea not to take any control measure and waiting for developing proper techniques or drugs  prior launching any control program . Notwithstanding these facts , management of animal excreta may greatly help in controlling parasitic diseases. If we start collecting all cattle dung to fill our gobar gas plants or to convert it into vermin-compose , this will not only control parasitic diseases but will also improve rural economy.

At last, I once again emphasize the need of a National Institute of Parasitology in country for paying proper attention on the parasitic diseases ,which are causing not only mortality but also production losses thereby affecting adversely national economy. I am avoiding repetition  what I have already said during  my lecture of Dr SC Parija oration gold medal award  in 2012 at Indore and  which has been published  in  Para Sight  2 Volume 2 ,issue 1 of IAAVP            and is also posted on my blog www.indianschistosomiasis.blogspot.com

SUMMARY :  

Launching of control program for any animal  disease which is causing mortality and production losses is important for any country . Ironically , the government of India  has launched control programs for viral and bacterial diseases but there is no control program ,ever launched by any government against any parasitic disease of our domestic animals. Neither , there is any system of collecting data ,related to animal parasitic diseases, from our veterinary hospitals and veterinary diagnostic laboratories . Therefore, it becomes difficult to answer- why we need to control parasitic diseases, which diseases are given priorities , and how we can control  these parasitic diseases ?

When we come to the question of how we can control animal schistosomiasis in India , we observed that our diagnostic technique is not sensitive due to low egg production of Indian schistosomes and requires further research. Likewise, the most effective Praziquantel drug is not able to kill significant number of Indian schistosomes with 35-50% left over blood-flukes . To circumvent this situation, it will be prudent to find out hyper-parasites of schistosomes and other parasites which will selectively kill these without harming their hosts. However, it will be futile to wait for developing proper techniques. Instead , animal excreta may be collected at mass scale and may be used in gobar gas plants or for preparing vermin-compose which will not only reduce prevalence of  parasitic diseases but will also improve rural economy.  The author has again emphasized opening a national institute of parasitology so that a holistic view may be developed towards parasitic diseases ,existing in the country .

REFERENCES :

Agrawal, M.C. 1999. Schistosomosis : an underestimated problem in animals in South Asia. World Animal Review 92 : 55-57.

Agrawal,M.C. 2003. Epidemiology of fluke infections . In (ed Sood ML)  Helminthology in India .Inrtenational Book Distributors, Deharadun . Page 511—542

Agrawal MC.  2012.  Schistosomes and schistosomiasis in South Asia. Springer India Pvt Ltd , New Delhi.

Agrawal MC ,Singh KP, George J and Gupta S 2005. Niclosamide trials on Indoplanorbis existus and Lymnaea luteola under different conditions. J Parasit Dis 29: 53-58

Dutt SC 1980. Paramphistomes and paramphistomiasis of domestic animals of India. Punjab Agriculture University, Ludhiana . Page 162.

Rathore BS  1998 . An epidemiological study on buffalo morbidity and mortality based on four year observations on 18630 buffaloes maintained at 28 livestock farms in India. Indian J Comp Microbiol.  19 :43-49

Shames N,Agrawal MC and Rao KNP 2000. Chemotherapeutic efficacy of praziquantel and closantel in experimental porcine schistosomiasis. Indian J Anim Sci 70 : 797-800

WHO  1985. The control of schistosomiasis. Technical report series 728. World Health Organization ,Geneva Page 113     

A DISCUSSION ON DIAGNOSIS OF ANIMAL SCHISTOSOMIASIS IN SOUTH ASIA

This is the lecture to be delivered in the  WINTER SCHOOL on MOLECULAR BIOLOGICAL APPROACHES FOR DIAGNOSIS AND CONTROL OF PARASITIC DISEASES at IVRI , Izatnagar in December ,2013. I am posting it on my blog for wider circulation among  scientific community, who are interested in schistosomiasis: will be pleased to see their comments. 

  

                                                                        M C AGRAWAL

                           FORMER NATIONATIONAL FELLOW AND EMERITUS SCIENTIST

                                 COLLEGE OF VETERINARY SCIENCE AND A H , JABALPUR 482001

                        drmcagrawal@gmail.com    www.indianschistosomiasis .blogspot.com

(Present address : ¾ Datt Arcade Phase Three, South Civil Lines, Jabalpur 482001)

Those who are working on diagnosis of schistosomiasis are aware how vast is the subject and its impossible for anyone to cover the topic in one hour lecture. Therefore, it will not be my goal to cover the whole subject , rather I will discussion some pertinent points on the topic at this occasion. We have discussed in detail about the diagnosis of schistosomiasis – both parasitological and immuno-diagnosis in our recently launched book(Agrawal,2012)  ,Schistosomes and schistosomiasis in South Asia (ISBN  9788132205388 www.springer.com) and you may refer this book for all the detailed information and related discussion . One advice is to those scientists who are interested to work on any problem of schistosomiasis in south Asia is to study well all the concerned  literature on the subject (as it is too vast ) prior undertaking any work as this prior study will inform them about the  work already done and the existing problems; such exercise will avoid duplication of work and will help in understanding well the present problem related to south Asian schistosomes and schistosomiasis which are certainly different from African or East Asian schistosomiasis .

Generally we are working and  discussing methodology of one or other diagnostic technique , its sensitivity and specificity , but there are no discussions whether  we need to devote our time , energy and funds for developing that  specific diagnostic technique and what will be its utility in present scenario. I think such discussion is necessary and my this lecture will focus on this topic while discussing schistosome diagnosis.

Further, by definition- as per Oxford dictionary , diagnosis means  “ statement of  nature of a disease or other condition made after observing its signs and symptoms” . Thus strictly diagnosis was restricted to what we call clinical diagnosis and as you know with passage of time it has gone in the back ground and laboratory diagnosis has taken a central stage-so is the case with schistosomiasis .One reason for this is our realization of difficulty in identifying the disease on symptoms alone due to commonness of them  in many other ailments. Therefore, we cannot whisk away laboratory diagnosis and in fact this is the main topic while discussing schistosomiasis diagnosis.  To this , I have taken liberty to include identifying schistosome infection in snail host ,as well- this is to understand the subject and related problems quite well.  

OUR AIM OF DIAGNOSIS : The basic question is what is our aim of identifying schistosome infection ? Whether it is only for treating a particular animal coming to veterinary hospital ? Or to declare a geographical area endemic with a given schistosome species ? Or to understand epizootiology of the infection ? Or to study zoonotic aspect of a schistosome species ? Or to know percentage of infected animals in each species ,age and sex wise ? Or to study immune status of animals after vaccination program ( so far ,no commercial vaccine is used against schistosomiasis in India ; it may be only on experimental basis ) . Or we wish to collect schistosomes during necropsy or in experimental animals for molecular and other studies  . Again schistosomiasis is existing in animals in two important forms (nasal schistosomiasis or snoring disease and hepato-intestinal schistosomiasis ) and diagnosis of both are different . So, there are varied aims of diagnosis hence our methodology of diagnosis should also differ as per our aim . Therefore, I will discuss the subject as per our aims and what best methodology we should follow to attain our aims of studies.

SENSITIVITY AND SPECIFICITY OF TESTS :

While starting the discussion, let me clarify that diagnosis of schistosomiasis is difficult due to two reasons . First, the symptoms in hepatic form like diarrhea, dysentery, anorexia, weakness, bottle jaw  etc are also present in other chronic or sub-acute infections . However, in snoring disease , the animal exhibits pathognomic signs like snoring sound, nasal discharge and nasal granuloma but totally depending on these symptoms may miss the infection in buffaloes or some cattle which are passing eggs in nasal discharge but without symptoms; and by this strategy , you cannot identify nasal schistosomiasis ,as is existing in Jabalpur , where local cattle are resistant to Schistosoma nasale but buffaloes are harboring the parasite- this  was confirmed by Banerjee and Agrawal (1991).

Therefore, for confirming the infection, we have to depend on laboratory diagnosis which is of two types –Parasitological and immunological- and both are having their own problems which have been discussed in chapter 7 and 8,respectively in above book. Basically , laboratory diagnosis is difficult due to poor sensitivity with no ante-mortem parasitological method being capable to correctly estimate schistosome prevalence in a given geographical area . For these reasons, I have argued about under estimation of schistosomiasis in whole of south Asia ( Agrawal, 1999)

 OUR PROBLEMS IN PARASITOLOGICAL METHODS

In parasitological diagnostic method , we can deal  only with  faeces (in hepatic form)  of the animal during ante-mortem  period . The most common method of faecal examination, followed in any veterinary diagnostic laboratory, is direct wet smear method . In some advance laboratories  sugar flotation or salt concentration method is also followed. Ideally , both supernatant and sediment should be examined for checking nematode and fluke infections but many times only supernatant is examined without caring for the sediment.  Ironically , examination of faeces by both these methods have resulted in identifying only 2-8% infection in any animal species whereas ,in reality , the infection rate was varying between 60-80% . Obviously, such diagnostic method cannot be recommended for diagnosing schistosomiasis when surveillance of the infection is our aim.

ANTEMORTEM DIAGNOSIS

Therefore, my laboratory assessed different coprological methods which may be used preferentially under field conditions. We have tried and compared direct smear, salt floatation,  acid-ether or formal-ether, sieving method, and Kato-katz methods in experimentally infected laboratory animals as well as  in field animals (Agrawal 2000 , Gupta 2002 , Vohra 2005  ). The results varied as per animal species, age of animal and intensity of the infection but sensitivity of these tests ascended from  direct smear to salt floatation, kato-katz , formal ether  methods leading highest level to 16% .

As any of the above egg detection method could not provide a practical solution of diagnostic problem, we tried hatching method by taking advantage that the eggs of schistosome species contain fully developed  miracidium , ready to hatch ,soon after coming in contact with  fresh water . There are many factors which influence this hatching of eggs and have been discussed elsewhere ( Agrawal,2012).

In summary , we can say that hatching method proved always superior over any egg detection method hence this is the test which we recommend to be followed in  field as well as in  research  laboratories. When we studied the method more meticulously , it was found more effective than any egg detection method where number of schistosome eggs in the faecal material was low- this is the young or old animals where egg concentration is low. For instance ,the hatching method ,though showed higher prevalence in pigs in comparison to egg detection methods, could not show significant variation in prevalence rate of porcine schistosomiasis at Jabalpur .This was because of higher faecal  concentration of schistosome eggs   in pigs (in comparison to other animal species) resulting its identification by other egg detection methods, as well.

The other  animal species where we observed different results, while comparing miracidia detection vis a vis egg detection , was experimentally infected albino mouse. Here, acid-ether method was more simple and sensitive than hatching method which failed many times to detect miracidia in the faecal material. 

The hatching method is not only simple but also cost effective as it does not require any chemical or use of micro slides. Therefore, this is an ideal method that can be followed under field conditions with little efforts . 

POST MORTEM METHODS :

Even diagnosing of schistosomiasis is not easy while conducting post mortem of an animal in comparison to other helminthic infections. Thus a simple cut in the liver may yield liver flukes, opening of rumen reveals amphistomes while intestine lumen will cause recovery of many nematode species . And in all these organs, you will not get schistosomes declaring the animal negative for schistosome infection. I may substantiate my statement by one reference of Rathor (1998) who compiled postmortem reports of more than eighteen thousand buffaloes of 28 livestock farms of India; the buffaloes were harboring Fasciola and amphistome infections but no case of schistosomiasis was recorded in these post mortem reports . As intermediate hosts of  amphistomes and schistosomes are identical ,it is difficult to believe that all the animals were negative for schistosomes-rather this is a case of failure of diagnosing schistosomiasis in any of these animals due to following routine post mortem techniques.  

This issue was attended by us and we have described specific techniques which should be followed during conducting post mortem of an animal for diagnosing schistosomiasis in them . These are two techniques ; one is perfusion technique for recovering alive blood flukes and the other is egg detection method using intestinal scrapings or liver pieces (Agrawal 2012, Vohra 2005  )    

IMMUNO-DIAGNOSIS :

A lot of research work  globally is being carried out on immuno-diagnosis mainly to study  human schistosomiasis in endemic countries and there are various reviews over the subject. It is difficult to cover them in one hour lecture . Nevertheless, I will mention some important facts which will help our discussion . For more details , you may read chapter 8 of the above referred book (Agrawal,2012).

ANTIBODY DETECTION METHODS :

In the beginning of the work, all the efforts were made to detect specific schistosome antibodies, developed by the host  due to schistosome pre- infection by using different types of the antigens .

There was fascination to use live larval stages of the blood flukes ,all three forms- viable eggs, miracidia, and cercariae- as experimental works have shown development of precipitates or blebs around viable schistosome eggs or immobilization of miracidia  or development of a hyaline membrane around cercariae when these larvae were immersed in the serum containing schistosome antibodies. The former two larvae were discarded as the tests were not very sensitive while CHR (cercarian hullen reaction ) proved a promising immunodiagnostic test though with the requirement of procuring alive cercariae for conducting the immunodiagnostic test.

To avoid the requirement of alive larvae, soluble antigens were developed using different methods and different stages of the schistosomes. Those who dealt the subject realized how difficult it is to recover a good quantity of schistosomes comparatively due to its smaller size and its location  in the blood vessels . Therefore, soon ,the immuno-diagnostic tests requiring larger quantity of antigens i.e. ring precipitation test were discarded . And in recent years ,all attention is diverted on ELISA ; as plate ELISA requires larger quantity of antigen and is more tedious , Dot ELISA is the method of choice among schistosomologists as each test requires only nano gram of antigens and is more simple to perform .

Looking to importance of diagnosis of fluke infections ,a NATP project was run by ICAR   with main center at  IVRI during 2000-2004 and  making Jabalpur, Hisar, as investigating centers for diagnosis of schistosomiasis ; some important work was also carried out at Bangalore veterinary college. Though these centers were able to develop Dot ELISA tests for diagnosing schistosomiasis with higher sensitivity , specificity could not reach to higher levels . Even some scientists ( Sumanth et al 2003) found  Dot ELISA positive in all the field animals who ever have been exposed to schistosome infections (?) and presently were either  negative or positive for schistosome eggs. What will be the utility of such diagnostic test in studying schistosomiasis remains debatable (Agrawal 2012).

ANTIGEN DETECTION METHOD :

The antibody detection methods have now been discarded globally by scientific community due to four  reasons. First , it does not differentiate between present and past infection . Second, it fails to calibrate intensity of the infection ; neither it reflects that all these positive animals are immune to re-infection . Third, it is difficult to collect blood from large animals under field conditions where controlling of the animals is a great problem (now there are ethical questions for collecting human blood merely for surveillance work). Fourth,the invasive method (collecting blood) merely for diagnosis is not favored due to the chances of transmitting viral and other infections; this has become more controversial in human cases where there are chances of transmitting HIV due to minor negligence of the technician.

Therefore, in recent years , attention has been diverted to detect schistosome antigen in faeces or urine of man and animals and principles of ELISA are followed for developing these methods. To make diagnostic test more specific and sensitive monoclonal antibodies have been used to attach them on nitrocellulose strip. Again , leading to  field adaptation of these tests , single step diagnostic kits have been developed where a solution is to pour on the strip which will develop two bands in case of positivity and only control band if the case is negative for schistosome infection.

Let me clarify that no commercial single step diagnostic kit is developed or available in India , though needed at least for diagnosing migratory human cases . This has been developed in some other countries  and one single step dipstick kit from European Veterinary Laboratory , Netherland was tried by us resulting in five positive human cases who were either suffering or were having history of cercarial dermatitis.

SCHISSTOSOMA SPECIFIC TESTS :

As this is the winter course on molecular studies on diagnosis and control of parasitic infections,  an important clarification , regarding all immuno-diagnostic methods is that they are capable to diagnose Schistosoma infection- only up-to genus level and none of these tests are species specific . Therefore, it is important to note that none of these tests will   inform which schistosome species is present in positive host species (in one sense , the test is good being able to be positive if the animal is infected by any schistosome species thus avoiding checking with many species specific tests- if and when developed  ). When it fails to identify schistosome species, any of these tests are of no use in identifying strains or sub-species of the schistosomes; neither they may help in detecting any new schistosome species.

Not only immuno-diagnostic methods, but our egg detection methods may fail to differentiate among schistosome species where egg morphology is closely resembling. If you study the literature, you will notice  that all  oval shaped schistosome eggs could not be ascribed to S.haematobium or S.indicum ; likewise , spindle shaped eggs does not mean only presence of S.spindale in the area ( Rollinson and Southgate 1987).

Therefore, now we have come to our main discussion –what is the aim of our diagnosis ?

It would have been clear from above discussion that the above methodology will not solve all our problems related to schistosomiasis and we have to stitch specific methods of diagnosis as per our requirements. I am discussing some of them in brief :

DIAGNOSIS FOR TREATMENT : When an animal with certain symptoms visit to a veterinary hospital , the main aim is the treatment of that particular animal. As the symptoms of schistosomiasis are common with other infections, though treatment is not the same, the differential diagnosis for schistosomiasis is important. The parasitological diagnostic tests have been termed as ‘Golden tests’  hence it is always recommended to diagnose these animals by coprological methods. This should include acid-ether or formal –ether method along with hatching method .The tests may be repeated twice or thrice if one time method shows negativity .

COLLECTION OF SCHISTOSOMES : If there is the need of collecting alive schistosomes from any animal species, perfusion method is the ideal one (Agrawal 2012). Recovery of schistosomes will also confirm presence of the infection in that particular host species and the geographical area to which the animal belonged  . In old days, the scientists teased the blood vessels for checking the animal positive for schistosomiasis and collecting alive blood – flukes; this method is tedious as well as less sensitive hence not recommended. Previously, it was difficult to apply perfusion technique as it was carried out by the use of a peristaltic pump which was costly and not available in Indian laboratories . however, we have replaced it  ,successfully, with a vertical water pump, most commonly used in desert coolers in Northern India (costing Rs 500/) hence now it is feasible to any laboratory to perform perfusion technique for recovering schistosomes during necropsy of the animals and the technique has been described in details in the book ( Agrawal,2012).

The alternative method, though less sensitive , is to collect mesentery or nasal cavity (when working on nasal schistosomiasis)  of the suspected animal, cut into small pieces and soak in Luke warm saline for 4-6 hours. Afterwards, the saline is filtered using a muslin cloth  which is transferred to  a petri-dish to collect the blood flukes ( Agrawal,2012).

IDENTIFYING NEW ENDEMIC AREA :  As you have come from different places of the country, it may be interesting for you to confirm whether a geographical area of your state or district or Tahseel is positive for any schistosome species. For confirming this possibility, you have to examine both intermediate and final hosts- in all seasons and in good number  else your results will not be reliable.

Till now only Indoplanorbis exustus and Lymnaea luteola have been identified as intermediate hosts for all the five well studied schistosome species i.e. Schistosoma indicum,S.spindale,S.nasale,S.incognitum and Orientobilharzia dattai while the snail host of Bivitellobilharzia nairi , O.bomfordi (?), S.haematobium (?- Gimvi infection) have still not been confirmed. Therefore, all the fresh water snails of the area should be examined meticulously for shedding of brevifurcated , apharyngeal ,non-ocillate  cercariae ; presence of such type cercariae will indicate prevalence of any mammalian schistosome species in the area (Agrawal,2012) .If these cercariae are shed by a new snail species, there are all chances that you are dealing with a new schistosome species .

As morphological studies of these cercariae cannot differentiate different mammalian schistosome species and certainly cannot confirm strains or sub-species of the schistosomes ,it is recommended to develop molecular techniques to study  these cercariae. So far, it is suggested that the snails are infected only with mono-species of fluke infections and molecular studies will be able to confirm or otherwise the above notion .

When you are examining final hosts for schistosome infection, first check if the animal is not a migratory animal otherwise origin of the infection will remain questionable. Therefore, area of birth of the animal is important in such studies and only by this methodology we suspected that Jabalpur is also harboring Schistosoma nasale in the buffaloes ( Banerjee and Agrawal 1991).

Though immuno-diagnostic methods are having high sensitivity (80-95%), their specificity ranks between 60-70% therefore solely depending on immuno-diagnosis while confirming a new endemic area is not a very good idea. However, not utilizing any immuno-diagnostic method may not reveal schistosome infection in a geographical area hence its use is important while exploring possibility of schistosomiasis in any animal species.  It must also be associated with sensitive parasitological methods which should always include hatching method. While examining animals for S.nasale ,instead of nasal discharge ,check the nasal scrapings as the latter is four  times more sensitive than the former method .

STUDYING ZOONOTIC POTENTIALS OF SCHISTOSOME SPECIES :  This is a topic of great interest in whole of South Asia but has not been studied meticulously . Identifying an endemic focus of urinary schistosomiasis in Gimvi village of Ratnagiri district, Maharashtra state ( Gadgil and Shah,1952) generated considerable interest in human schistosomiasis in the country but non –detection of clinical urinary cases from other parts of the country created a complacent view of absence of human schistosomiasis in other parts of the country . This view was developed ignoring the fact that S.incognitum was first detected in two human cases by Chandler (1926) and there are reports of finding schistosome eggs in human excreta in other parts of the country  (Agrawal,2012) . The topic has been discussed in more details while reviewing present status of schistosomiasis in India ( Agrawal,2005).  Prevalence of cercarial dermatitis in rural India is very common (Agrawal,2012) although no further studies have been carried out on the ailment. Thus we are not sure to what stage of development the schistosomula reaches in humans showing different immunological conditions  .

In the above cases, generally parasitological methods have failed to detect schistosome eggs in human excreta ,therefore, it is advocated to use a more sensitive and specific antigen detection test . The specificity of single step diagnostic kits , developed by using S.mansoni or S.haematobium, in other countries is not very high warranting developing such kits using Indian schistosome species.  Likewise, we do not have any specific immunological test for confirming cercarial dermatitis cases ; this is only history of dermatitis  and recovering schistosome cercariae from the fresh water snails which is collaborated with the ailment. Therefore, it will be beneficial to develop an immunodiagnostic test which may confirm cercarial dermatitis- ideally with possibility of differentiating cercarial dermatitis caused by avian schistosome cercariae and that by mammalian schistosome cercariae.

As you will notice that the single step diagnostic kit or immunodiagnostic test for cercarial dermatitis will not be able to identify schistosome species or strains involved in each case. Therefore, there is the need to develop molecular techniques for identifying species or hybrids or strains of schistosomes ,existing in whole of south Asia. Obviously, both the methodologies are supplementary to each one in solving the problem of schistosomiasis in the region.

CHECKING HOST  IMMUNITY AGAINST SCHISTOSOMES :  In viral and bacterial infections , immunological tests have also been used to check immune status of a given population and are good tools for vaccination programs . When we look the scenario with relation to schistosomiasis , two facts have emerged .

First, there is no vaccine which is being used for controlling schistosomiasis . This is the case both for human schistosomiasis as well as for animal schistosomiasis . More and more efforts are being made to develop a vaccine which may protect the host against schistosome infection. I am not aware if any institute in India is trying to develop a feasible vaccine for schistosome infection in any host species.

Secondly, our immuno-diagnostic tests are able to reveal presence of antibodies in positive cases but whether presence of these particular antibodies   will be able to protect the host against re-infection with schistosomes is questionable .So far , we have not developed any immuno-diagnostic test  whose positivity also guarantee protection of the host against re-infection.

SURVEILLANCE OF ANIMALS FOR SCHISTOSOMIASIS : Whenever , we undertake surveillance work for identifying any infection, our diagnostic test should be most sensitive and specific in diagnosing that infection. Ironically, no diagnostic test stands on above criteria with regards to schistosomiasis  . With Indian schistosomes , parasitological tests are least sensitive due to comparatively  lower egg excretion  hence their sole use in surveillance work is never advocated . In our opinion, hatching method has proved  most sensitive parasitological  method during ante-mortem hence must be included in all surveillance works. Those cases, positive by hatching method, must also be examined by a more sensitive egg detection method which will help  in identifying schistosome species of that area ,as well.

Since parasitological tests are least sensitive, the surveillance work must be carried out by supplementing parasitological methods with a more sensitive and specific immuno-diagnostic method. All the work , carried out in India, have proved Dot ELISA as most promising immuno-diagnostic test hence this should be incorporated in surveillance work but fully understanding its limitations (Agrawal,2012).

The antigen detection test in animal faeces will be more simple and be able to detect existing schistosome positive cases hence there is the need to develop these tests for understanding schistosomiasis in South Asia.

As we have already discussed the limitations of these immuno-diagnostic tests as well as those of parasitological tests, schistosomes and schistosomiasis cannot be understood well in whole of south Asia without developing more sophisticated molecular techniques which will help in identifying new schistosome species and also hybrids of the schistosomes which have developed in the continent due to hybridizations with existence of two or more schistosome species in a single host species. A collaboration with some international laboratories who are working on these topics since long  will make the efforts more easy ,accurate ,providing the results in shorter time frame .  

SUMMARY : The symptoms in hepatic schistosomiasis are vague hence laboratory diagnosis is carried out for confirmation of the disease. While deciding to employ diagnostic methodology, it is important to consider our aim of diagnosing the infection. When our sole aim is treating a sick animal, visiting a Veterinary Hospital ,coprological diagnosis is recommended but using more sensitive methods like acid-ether and hatching methods. During post-mortem ,either for collecting blood –flukes or for confirming the infection, perfusion method is the choice having highest sensitivity . In deciding zoonotic potentials of a given schistosome species or for surveillance of the infection  both parasitological and immunological methods should be followed as both will supplement each other. As antibody detection method does not differentiate between present and past infection with additional disadvantage of invasiveness, antigen detection methods in host’s excreta are gaining priorities .But it must be remembered that so far all immunodiagnostic methods are efficient only to identify the infection up-to genus level ( Schistosoma )  and cannot differentiate the infection  up-to species levels . Therefore, it is essential to undertake molecular  studies , if you wish to decide existence of schistosome species, sub-species or hybrids , in a given geographical area,   as no immuno-diagnostic method can reveal such complexities of schistosomiasis .

REFERENCES :

Agrawal, M.C. 1999. Schistosomosis : an underestimated problem in animals in South Asia. World Animal Review 92 : 55-57.

Agrawal, M.C. 2000. Final report on National Fellow project “Studies on strain identification,epidemiology,diagnosis,chemotherapy and zoonotic potentials of Indian schistosomes. ICAR, New Delhi

Agrawal,M.C. 2005. Present status of schistosomosis in India. Proc Nat Acad Sc, India. 75 (B),Special issue : 184-196

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Chandler AC.  1926.  A new schistosome infection of man with note on other human fluke infections in India. Indian J Med Res 14 : 179-183

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