Molecular characterization and histo-physiological alterations induced by concurrent helminthosis in the liver of urban commensal rodents in Punjab, India.

BACKGROUND: Rodents harbour a number of parasites of public health importance, thus, they threaten human health and livestock. AIMS: The present study aimed to characterize two helminthic species found in commensal rodents and record histo-physiological alterations induced by them. METHODS: A total of 300 synanthropic rodents of three species: Rattus rattus (n=201), Bandicota bengalensis (n=90), and Mus musculus ( n=09 ) were live trapped and necropsied in different seasons during November 2017 to October 2019 at Ludhiana, Punjab, India. RESULTS: Liver of two species B. bengalensis (72.22%) and R. rattus (65.67%) were found infected with two helminthic parasites Taenia taeniaeformis, and Calodium hepaticum. These endoparasites were present either alone (4.33-6.33%) or as mixed infection (65.55%). The level of total proteins and liver marker enzymes including aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were found significantly higher in the liver of rodent species infected with single and mixed infection compared to those with no infection. In histopathological assay, granulomatous liver lesions and necrosis of hepatocytes were seen which were associated with eggs and adults of C. hepaticum and inflammatory reaction in hepatic parenchyma adjoining to cysts of T. taeniaeformis. Based upon scanning electron microscopy (SEM) identification and molecular characterization using mitochondrial cytochrome oxidase I (COI) region, the metacestodes in whitish cysts were confirmed to be of T. taeniaeformis for the first time in Punjab, India. CONCLUSION: The study highlights an alarmingly high infection of rodents with zoonotic parasites and suggests immediate pest (rodent) control to check the dissemination of zoonotic diseases by helminth species under study.

Atypical theileriosis with cutaneous involvement in a cow in India: a case report.

Bovine tropical theileriosis caused by Theileria annulata is an overwhelming haemoprotozoan tick-borne disease in taurine and cross-bred cattle in Punjab, India. However, there seems to be no report from India of cutaneous nodules associated with the disease. This report describes a five-year-old cross-bred cow presented to a university clinic with a history of fever, inappetence and malaise for the past six to seven days. Clinical examination revealed normal vital parameters, pale mucous membranes, mild enlargement of the prescapular lymph nodes and multiple subcutaneous nodular masses (2-4 cm) on the neck and abdomen. Haematology revealed mild anaemia and leucopenia with 48% neutrophils, 48% lymphocytes and 4% eosinophils. Romanowsky-stained smears of fineneedle aspiration biopsy samples from swollen lymph nodes and subcutaneous masses showed an increased number of lymphoid cells, suggesting cutaneous lymphomatosis. However, a critical examination of the smears from subcutaneous nodules showed a large number of Koch's blue bodies in macrophages and lymphoblasts, and several piroplasms were also noticed within the red blood cells in lymph node smears. A peripheral blood smear revealed mild to moderate parasitaemia. Extracted DNA from the parasitologically positive blood sample was subjected to nested polymerase chain reaction (nPCR) using T. annulata speciesspecific primers encoding the 30-kiloDalton major sporozoite surface antigen. The desired 572-base pair amplified product of the nPCR was comparable to the positive control. This seems to be a rare case of T. annulata in an adult cross-bred cow, showing cutaneous nodular involvement.

La theilériose bovine tropicale est une maladie causée par le protozoaire Theileria annulata et transmise par les tiques, affectant massivement les populations de bovins et de bovidés métis au Pendjab (Inde). Il semble toutefois que la présence de nodules cutanés associés à la maladie n'y ait jamais été rapportée jusqu'à présent. Les auteurs décrivent le cas soumis à une clinique vétérinaire universitaire d'une vache métisse âgée de cinq ans qui présentait depuis six à sept jours un tableau fébrile accompagné d'une perte d'appétit et d'un affaiblissement général. À l'examen clinique, les paramètres vitaux étaient normaux mais une pâleur des membranes muqueuses a été observée, ainsi qu'un gonflement modéré des ganglions lymphatiques préscapulaires et de nombreuses masses nodulaires sous-cutanées (de 2 à 4 cm d'épaisseur) au niveau du cou et de l'abdomen. L'hématologie a mis en évidence une anémie modérée et une leucopénie, les leucocytes se répartissant en 48 % de neutrophiles, 48 % de lymphocytes et 4 % d'éosinophiles. Les frottis à coloration de Romanowsky d'une biopsie par aspiration à l'aiguille fine des ganglions lymphatiques enflés et des masses sous-cutanées ont fait apparaître une augmentation du nombre de cellules lymphatiques évocatrice d'une lymphomatose cutanée. Néanmoins, un examen critique des prélèvements de nodules sous-cutanés a permis de constater la présence d'un grand nombre de corps bleus de Koch dans les macrophages et les lymphoblastes ; en outre, de nombreux piroplasmes ont été trouvés dans les globules rouges des frottis de ganglions lymphatiques. Un frottis de sang périphérique a permis de quantifier la parasitémie comme étant de niveau faible à modéré. L'ADN extrait de l'échantillon de sang à parasitologie positive a été soumis à une amplification en chaîne par polymérase nichée (nPCR) utilisant des amorces spécifiques de T. annulata codant pour l'antigène majeur de surface (30 kDa) du sporozoïte. Le produit amplifié par nPCR de la séquence souhaitée de 572 paires de bases était similaire à celui de l'échantillon de contrôle positif. Il s'agit probablement d'un cas rare d'infection à T. annulata chez une vache adulte métisse présentant des manifestations nodulaires cutanées.

La teileriosis tropical bovina causada por Theileria annulata es una devastadora enfermedad hemoprotozoaria transmitida por garrapatas que afecta al ganado taurino e híbrido del Punjab (India). Ahora bien, en la India no parece haber ningún caso descrito de esta enfermedad que se acompañe de la presencia de nódulos cutáneos. Los autores describen el caso de una vaca de cinco años híbrida que fue presentado a una clínica universitaria con un cuadro de fiebre, pérdida de apetito y decaimiento en los seis a siete días anteriores. El examen clínico puso de manifiesto parámetros vitales normales, mucosas pálidas, leve hipertrofia de los ganglios linfáticos prescapulares y múltiples bultos subcutáneos de tipo nodular (2 a 4 cm) en cuello y abdomen. El análisis hematológico reveló una leve anemia y leucocitopenia, con un 48% de neutrófilos, un 48% de linfocitos y un 4% de eosinófilos. Tras proceder a una biopsia de ganglios inflamados y bultos subcutáneos por aspiración con aguja fina, el examen de frotis de estas muestras con tinción de Romanowsky reveló un número excesivo de células linfáticas, lo que parece apuntar a una linfomatosis cutánea. No obstante, al examinar más a fondo los frotis de nódulos subcutáneos se observó que macrófagos y linfoblastos albergaban un gran número de cuerpos azules de Koch. También se observaron varios piroplasmas dentro de los eritrocitos presentes en los frotis de ganglios linfáticos. Un frotis de sangre periférica reveló una parasitemia entre leve y moderada. El ADN extraído de esta muestra de sangre positiva fue sometido a una técnica de reacción en cadena de la polimerasa (PCR) anidada en la que se emplearon cebadores específicos de la especie T. annulata que codifican el antígeno de superficie principal de 30 kDa del esporozoíto. La deseada secuencia de 572 pares de bases amplificada por PCR resultó comparable con la correspondiente secuencia de la muestra positiva de control. Parece tratarse pues de un caso raro de infestación por T. annulata de una vaca adulta híbrida que se acompaña de nódulos cutáneos.

Patho-physiological observations in natural concurrent infections of helminth parasites of zoonotic importance in the wild rodents, Bandicota bengalensis.

Screening of 83 mature wild rodents of Bandicota bengalensis (52 male and 31 females), live trapped from premises near railway station, fish market and agricultural fields in Ludhiana district of Punjab province, India was carried out to assess the natural occurrence of helminth parasites and their potential as reservoirs of zoonoses. On necropsy, intestine of rats was found infected with adult cestode parasites of Hymenolepis diminuta and H. nana. Liver was found infected with Capillaria hepatica and Cysticercus fasciolaris. These parasites were present either alone or as concurrent infection. In overall, 68.67 % rats were found infected of which 59.65 % were infected with nematode parasite, C. hepatica and 75.44 % with cestode parasites. Grossly, pathognomonic lesions of C. hepatica and C. fasciolaris were seen clearly and microscopic changes were observed only in severely infected cases. Overall investigation on the effect of these parasitic infections on pathophysiology of the host revealed that rodents serve as reservoirs of these parasitic infections without having much pathogenic effect on their vital organs, fecundity and enzyme activities. Because of its close association with human habitations, B. bengalensis may act as an important source of zoonotic infections and thus direct or indirect contact with excrements and carcasses of this rodent species, should be avoided.

Induced neurocysticercosis in rhesus monkeys (Macaca mulatta) produces clinical signs and lesions similar to natural disease in man.

Neurocysticercosis is a serious endemic zoonosis resulting in increased cases of seizure and epilepsy in humans. The genesis of clinical manifestations of the disease through experimental animal models is poorly exploited. The monkeys may prove useful for the purpose due to their behavior and cognitive responses mimicking man. In this study, neurocysticercosis was induced in two rhesus monkeys each with 12,000 and 6,000 eggs, whereas three monkeys were given placebo. The monkeys given higher dose developed hyperexcitability, epileptic seizures, muscular tremors, digital cramps at 10 DPI, and finally paralysis of limbs, followed by death on 67 DPI, whereas the monkeys given lower dose showed delayed and milder clinical signs. On necropsy, all the infected monkeys showed numerous cysticerci in the brain. Histopathologically, heavily infected monkeys revealed liquefactive necrosis and formation of irregular cystic cavities lined by atrophied parenchymal septa with remnants of neuropil of the cerebrum. In contrast, the monkeys infected with lower dose showed formation of typical foreign body granulomas characterized by central liquefaction surrounded by chronic inflammatory response. It was concluded that the inflammatory and immune response exerted by the host against cysticerci, in turn, led to histopathological lesions and the resultant clinical signs thereof.

Pathological alterations in natural cases of Capillaria hepatica infection alone and in concurrence with Cysticercus fasciolaris in Bandicota bengalensis.

The present study reports the natural occurrence and pathomorphological alterations of Capillaria hepatica infection alone and in concurrence with Cysticercus fasciolaris infection in the liver of Bandicota bengalensis. Out of the eighteen mature male B. bengalensis autopsied, livers of eight rats (44.4 %) were found infected with parasites comprising two (11.1 %) rats infected with C. hepatica alone, four (22.2 %) infected with C. fasciolaris alone and two (11.1 %) infected with C. hepatica in concurrence with C. fasciolaris. Gross lesions comprising of pale cystic areas or streaks on the surface of liver in rats revealed the presence of eggs of C. hepatica scattered in the parenchyma of the liver. Histologically, granulomatous reaction around the eggs, adult worms and dead components of parasites were observed.

Cytopathology of parasitic dermatitis in dogs.

Out of 44 cases of dermatitis in dogs, 11 cases of parasitic origin were analyzed by cytopathology. Histopathologic examination of punch biopsies was also done for correlation with cytologic findings. Sarcoptic dermatitis was recorded in six cases, wherein, besides sarcoptic mites, neutrophils, macrophages, and plasma cells and keratinizing epithelial cells were also seen. Hematology revealed a relative neutrophilia and mild eosinophilia. Four cases of severe and generalized demodicosis complicated with bacteria and/or Malassezia sp. infection were also recorded. Histopathologically numerous Demodex sp. mites in varying stage of maturation were found damaging the hair follicles along with associated pathological changes and foreign body granulomas in one case. In addition, flea allergy dermatitis was also observed in one dog. In nutshell, cytology was found to be unequivocally effective in diagnosing parasitic dermatitis.