Increased Fas ligand expression of peripheral B-1 cells correlated with CD4+ T-cell apoptosis in filarial-infected patients.

Cellular hyporesponsiveness observed during helminth infections is attributed to factors such as antigen-presenting cells (APC) dysfunction, increased interleukin-10(IL-10), regulatory T cells and induction of CD4+ T (Th)-cell apoptosis. Increased Fas ligand (FasL) expression on the surface of B-1 cells and induction of apoptosis of Th cells by FasL-expressing B-1 cells due to helminth infection were demonstrated in murine model of helminth infection where as profile of FasL expression, Th-cell apoptosis and correlation between these two populations of cells in clinical filariasis remain unknown. In this study, we have scored the profile of apoptotic Th-cell population and FasL-expressing B-1 cells in different clinical categories of filariasis. The peripheral apoptotic T-helper cells were significantly increased in filarial patients compared to endemic controls. Expression of FasL on the surface of peripheral B-1 cells increased in filarial patients and positively correlated with peripheral apoptotic T-helper cells indicating FasL-expressing B-1 cells may be one of the important mediators of Th-cell apoptosis and immune anergy during filarial pathology.

Comparative seroprevalence and risk factor analysis of Trypanosoma evansi infection in equines from different agro-climatic zones of Punjab (India).

As parasitaemia is low and fluctuating during the chronic stage of infection, accurate detection of Trypanosoma evansi in blood is difficult. The primary aims of this investigation were to assess for the first time the seroprevalence of T. evansi in all agro-climatic zones of Punjab, by indirect enzyme-linked immunosorbent assay (iELISA) and card agglutination test (CATT/T. evansi), and to evaluate the risk factors associated with latent trypanosomosis. A total of 319 equine serum samples collected from 12 districts of Punjab (India) belonging to different agro-climatic zones revealed 39 (12.23%) and 9 (2.82%) samples to be positive by CATT/T. evansi and iELISA, respectively. The highest prevalence was recorded from the Ludhiana district (42.86% and 7.14% by CATT/T. evansi and iELISA, respectively) in the central plain zone (for which the overall prevalence was 15% and 4.17%, respectively). There was fair agreement between the tests for the detection of T. evansi (kappa = 0.345). Species was the most influential risk factor for infection, with odds ratios (ORs) of 2.81 and 5.63 for donkeys/ mules, in comparison with horses, by CATT/T. evansi and iELISA, respectively. The female equine population (OR = 3.13, 95% confidence interval [CI] = 1.32-7.67 [CATT]) was found to be at a higher risk of seropositivity for T. evansi, particularly on 'unorganised' (inappropriately managed) farms (OR = 3.18, 95% CI = 1.53- 6.65 [CATT]) and among animals used for commercial purposes (OR = 2.51, 95% CI = 1.20-5.21 [CATT]). In conclusion, to declare disease-free status, use of the iELISA followed by retesting of suspect samples by CATT/T. evansi is suggested.

La détection minutieuse de Trypanosoma evansi dans le sang est difficile en raison du nombre faible et fluctuant de parasites pendant la phase chronique de l'infection. L'étude présentée par les auteurs vise, d'une part, à réaliser une première évaluation de la prévalence sérologique de T. evansi dans chacune des zones agro-climatiques du Pendjab en utilisant une épreuve immuno-enzymatique (ELISA) indirecte et le test d'agglutination sur carte pour la trypanosomose (CATT/T. evansi) et, d'autre part, à évaluer les facteurs de risque associés à une présence inapparente de la trypanosomose. Au total, sur les 319 sérums d'équidés prélevés dans 12 districts du Pendjab (Inde) appartenant à des zones agro-climatiques différentes, 39 échantillons (12,23 %) ont donné des résultats positifs avec le CATT/T. evansi et 9 échantillons (2,82 %) ont donné des résultats positifs à l'ELISA indirecte. La prévalence la plus élevée a été enregistrée dans le district de Ludhiana (42,86 % de résultats positifs avec le CATT/T. evansi et 7,14 % de résultats positifs avec l'ELISA indirecte) dans la zone des plaines centrales (où la prévalence globale s'élevait, suivant les méthodes de test, à 15 % et 4,17 %, respectivement). La détection de T. evansi par les deux tests a été concordante (kappa = 0,345). Le facteur de risque ayant le plus d'influence sur la probabilité d'infection était l'espèce, ce risque étant plus élevé chez les ânes et les mulets que chez les chevaux (rapport de cotes [odds ratio, OR] de 2,81 [CATT/T. evansi] et de 5,63 [ELISA indirecte]). Les femelles présentaient également un risque plus élevé de posséder des anticorps anti-T. evansi que les mâles (OR = 3,13 ; intervalle de confiance [IC] à 95 % : 1,32­7,67 [CATT]), en particulier dans les élevages « informels ¼ (sans gestion sanitaire) (OR = 3,18 ; IC à 95 % : 1,53­6,65 [CATT]) ainsi que parmi les animaux utilisés à des fins commerciales (OR = 2,51 ; IC à 95 % : 1,20­5,21 [CATT]). En conclusion, pour la démonstration de l'absence d'anticorps, les auteurs recommandent d'utiliser l'ELISA indirecte puis de soumettre les échantillons douteux à un test de confirmation au moyen du CATT/T. evansi.

La detección precisa de Trypanosoma evansi en la sangre resulta difícil porque en la fase crónica de la infección la parasitemia es baja y fluctuante. Los autores describen una investigación encaminada principalmente a determinar por primera vez la seroprevalencia de T. evansi en todas las zonas agroclimáticas del Punjab por ensayo inmunoenzimático indirecto (ELISAi) y por aglutinación en placa, así como los factores de riesgo asociados a la tripanosomosis latente. De un total de 319 muestras de suero equino procedentes de 12 distritos del Punjab (India) situados en diferentes zonas agroclimáticas, la aglutinación en placa deparó resultado positivo en 39 de ellas (un 12,23%) y el ELISAi en 9 (2,82%). El máximo nivel de prevalencia se registró en el distrito de Ludhiana (42,86% y 7,14% por aglutinación en placa y ELISAi, respectivamente), sito en la zona de la planicie central (que en conjunto deparó una prevalencia del 15% y el 4,17%, respectivamente). Ambas pruebas resultaron bastante coincidentes por lo que respecta a la detección de T. evansi (coeficiente kappa = 0,345). El factor de riesgo más influyente resultó ser la especie: en comparación con los caballos, los asnos o mulas presentaban una razón de probabilidad (RP) de 2,81 y 5,63 para la aglutinación en placa y el ELISAi respectivamente. Se observó que la población de yeguas (RP = 3,13; intervalo de confianza [IC] al 95% = 1,32­7,67 [aglutinación en placa]) presentaba un riesgo más elevado de seropositividad para T. evansi, especialmente en explotaciones «desorganizadas¼ (mal gestionadas) (RP = 3,18; IC 95% = 1,53­6,65 [aglutinación en placa]) y entre los animales utilizados con fines comerciales (RP = 2,51; IC 95% = 1,20­5,21 [aglutinación en placa]). Los autores concluyen proponiendo que, a los efectos de declarar la ausencia de enfermedad, se utilice en primer lugar el ELISAi, seguido de la prueba de aglutinación en placa para las muestras sospechosas.

Increased IgG antibody responses to excretory/secretory antigens in neonates born from mothers infected with filarial nematodes.

The present investigation aimed to evaluate the extent to which maternal filarial infection influences IgG subclass immune responses in the cord blood of neonates. Prevalence of antigenaemia was detected using an Og4C3 assay. Filaria-specific IgG subclasses against excretory/secretory antigens were measured by ELISA. Transplacental transfer of circulating filarial antigen (CFA) was observed from 34.8% of CFA-positive mothers to their respective cord bloods. Filaria-specific IgG1, IgG2 and IgG4 responses were significantly higher among cord bloods of infected mothers compared to cord bloods of uninfected mothers. In contrast, the IgG3 response was significantly higher among cord bloods of uninfected mothers. The study shows that transplacental transfer of filarial antigens and filaria-specific IgG4 occurs more in mothers having high worm burdens, and transfer of filaria-specific IgG3 occurs more in the cord blood of uninfected mothers. The findings of the study provide evidence for the development of prenatal sensitization to filarial antigens in utero, and high filaria-specific IgG4 in cord blood may serve as a marker for in-utero sensitization.

PCR and ELISA vis-à-vis microscopy for detection of bovine anaplasmosis: a study on associated risk of an upcoming problem in North India.

This investigation demonstrates the status of bovine anaplasmosis caused by A. marginale in bovines from Submountain and Undulating Zone of Punjab. Out of 184 suspected animals, 25 (19.51%), 47 (31.71%), and 78 (68.75%) were positive by microscopy, indirect ELISA, and PCR assay, respectively. The microscopy showed 29% sensitivity and 99% specificity, while ELISA showed 32% sensitivity and 79% specificity in concordance with PCR assay. Five false negative samples by msp1ß PCR were reconfirmed for Anaplasma spp. targeting 16S rRNA gene. The sequence analysis showed the presence for A. marginale specific restriction site, indicating variation in the local strains of the organism resulting in no amplification with msp1ß gene primers. Of 82 samples positive by PCR, 57 were negative by ELISA indicating lower efficacy of ELISA to detect early anaplasmosis. The assessment of risk factor with results of PCR technique indicated that cattle (Odds ratio = 2.884), particularly those of age > 1 years (Odds ratio = 2.204) of district Pathankot (Odds ratio = 3.182) of Submountain Zone (Odds ratio = 2.086), were at high risk of anaplasmosis. All three districts of Submountain Zone are at higher risk indicating the impact of biotic and abiotic factors on the incidence of disease.

In utero sensitization modulates IgG isotype, IFN-γ and IL-10 responses of neonates in bancroftian filariasis.

In utero exposure has been considered as a risk factor for filarial infection. To evaluate the influence of maternal infection on filarial-specific IgG subclass response in neonates and their correlation with plasma levels IL-10 and interferon-γ, 145 pairs of mothers and their respective cord bloods were examined. Transplacental transfer of circulating filarial antigen (CFA) was observed in 34·8% cord bloods from CFA positive mothers. Filarial-specific IgG1, IgG2 and IgG4 responses of cord bloods were found to be positively correlated with CFA of mothers. In contrast, IgG3 responses negatively correlated with CFA of mothers. The % of similarity of recognition pattern in the cord blood with maternal blood was high for IgG3 response than IgG4 in all three groups. An increased levels of IL-10 and decreased levels of interferon gamma (IFN-γ) were observed in cord blood of infected mothers. Interferon gamma was positively correlated with IgG3 and negatively correlated with IgG4 level. On the other hand, IL-10 was positively correlated with IgG4 and CFA, indicating that cytokines may play a role in modulating the immune responses in cord bloods of sensitized foetus. The findings of the study reveal that in utero tolerance or sensitization may influence the filarial-specific immunity to infection in neonates.

Hydrocele fluid: can it be used for immunodiagnosis of lymphatic filariasis?

BACKGROUND & OBJECTIVES: Diagnosis of lymphatic filariasis using serum has been established but the utility of hydrocele fluid for the purpose is not exactly known. Since, hydrocele is a chronic form of the disease manifestation in a variety of situations and often poses difficulty in diagnosing its origin, we have evaluated the usefulness usage of hydrocele fluid for diagnosis of filarial origin of hydrocele in this study. METHODS: Paired samples of serum and hydrocele fluid from 51 individuals with hydrocele, living in an endemic area of Wuchereria bancrofti were assessed. Circulating filarial antigen, filarial specific antibody and cytokine assay were performed in both serum and hydrocele fluid of patients. RESULTS: Og4C3 assay detected circulating filarial antigen (CFA) in serum and corresponding hydrocele fluids. The level of IgG, IFN-γ and IL-10 were found to be high in CFA-negative, while IgM and IgE were high in CFApositive hydrocele fluid and serum samples associated with hydrocele. On the other hand neither CFA-positive nor CFA-negative hydrocele fluid and serum samples associated with hydrocele showed any difference in IgG4 level. INTERPRETATION & CONCLUSION: This study showed that the filaria related antigens and antibodies found in serum can be detected with equal sensitivity in hydrocele fluid. Therefore, it can be used as an alternative to serum for immunodiagnosis of filariasis, and help monitoring the filarisis elimination programme.

Immuno-epidemiology of bancroftian filariasis: a 14-year follow-up study in Odisha, India.

Forty asymptomatic, circulating filarial antigen negative (CFA(-ve)) and ten asymptomatic, circulating filarial antigen positive (CFA(+ve)) individuals were followed up longitudinally over a period of 14 years at intervals of 7 years in order to investigate the immunological, parasitological and clinical changes that took place in an endemic area due to natural process. The clinical status, microfilaremia, circulating filarial antigenemia and immunological responses to filarial antigens (DSSd1 and Sd30) prepared from cattle filarial parasite Setaria digitata, were examined. The observations showed that 19 individuals had developed either antigenemia or filarial symptoms (acute filarial lymphangitis/hydrocele) from CFA(-ve) group. Three individuals had cleared antigenemia and one had developed microfilaremia from CFA(+ve) group after 7 years. Increased IgG and IgM and low IgG2 and IgG4 level responses along with high lymphocyte production were observed in CFA-negative individuals. This was in contrast to observations made in CFA(+ve) subjects. The results of the present study indicated that the changes taking place in the immunological, clinical and CFA status of individuals residing in filaria endemic regions developed different clinical manifestation with course of time.

Maternal filarial infection: association of anti-sheath antibody responses with plasma levels of IFN-γ and IL-10.

Maternal filarial infection influences the risk of acquiring infection and development of immunity in children. Here we have analysed the blood samples of 60 mothers (24 infected and 36 uninfected) and their corresponding cord bloods to assess the impact of maternal infection on the anti-sheath antibodies and cytokine production in neonates born from them. About 69·4% of non-infected mothers and their cord bloods showed the presence of anti-sheath antibodies, while only 16·6% of the cord bloods from infected mothers were positive for it. The IL-10 level was significantly high in cord bloods of infected mothers compared with non-infected mothers. At the same time the IL-10 level was also observed to be remarkably high in cord bloods of both infected and non-infected mothers negative for anti-sheath antibody. In contrast, IFN-γ levels were significantly high in cord bloods of non-infected mothers compared with infected mothers and the increment was prominent in cord bloods of both infected and non-infected mothers positive for anti-sheath antibody. The study reveals that the presence or absence of anti-sheath antibodies in association with cytokines skews the filarial specific immunity to either Th1 or Th2 responses in neonates. This may affect the natural history of filarial infection in early childhood.

Transplacental transfer of filarial antigens from Wuchereria bancrofti-infected mothers to their offspring.

OBJECTIVE: Maternal infection has been considered to be a risk factor for filarial infection in offspring. In order to examine the influence of maternal infection in neonates, we have determined the prevalence of circulating filarial antigen (CFA) and anti-filarial antibodies in 119 maternal and corresponding cord blood samples collected from an area endemic for bancroftian filariasis. METHOD: Prevalence of antigenaemia was detected using Og4C3 circulating filarial antigen enzyme-linked immunosorbent assay. The presence of microfilariae was determined by filtration of a 1 ml sample through a Nuclepore membrane. Antibody isotypes (IgG, IgM, and IgE) to filarial antigen (Setaria digitata antigenic extract) were determined by enzyme linked immunosorbent assay (ELISA). RESULTS: Microfilariae were detected in 14 cases (11.8%), whereas the Og4C3 assay could detect filarial antigen in 44.5% of pregnant mothers. Interestingly, 24.5% of samples born from CFA-positive mothers were found positive for CFA. None of the cord samples from CFA-negative mothers were found positive for CFA. No significant difference was observed in prevalence of filarial-specific IgG, IgM and IgE antibodies in CFA-positive and negative mothers. IgG antibody was detected in 60.5% of maternal and 21.8% of cord samples. IgG antibody in the cord does not differ with the antigen status of the mother. In contrast IgM and IgE antibody prevalence was significantly higher in cord from infected mothers than non-infected mothers (11.3% vs 0 for IgM, 24.5% vs 3.03% for IgE). CONCLUSION: Our study demonstrates the transplacental transfer of circulating filarial antigen from mother to cord. Filaria-specific IgM and IgE antibodies were higher in cord blood from infected mothers than from non-infected mothers. The findings of the study provide additional circumstantial evidence for pre-natal sensitization to filarial antigens developed in utero.

Effect of protein kinase C and protein kinase A inhibitors on contraction of isolated femoral arteries of SHR and Wistar rats.

Alterations of calcium handling and other second messenger cascades including protein kinase C (PKC) and A (PKA) were suggested to be responsible for abnormal vascular function in spontaneously hypertensive rats (SHR). However, the relative contribution of these pathways to vasoconstriction is still not completely understood. We investigated the effect of Ro 31-8220 (PKC inhibitor) and H89 (PKA inhibitor) on vasoconstriction induced by 120 mM KCl or by addition of 10 microM noradrenaline (NA) in isolated femoral arteries of control Wistar rats and SHR. Moreover, we investigated these responses in the presence and absence of Ca(2+) ions in the incubation medium in order to assess the role of calcium influx in these contractions. We observed that while the vasoconstriction in the presence of calcium was not different between Wistar and SHR, the difference between constriction elicited by NA addition in the absence and presence of external calcium was larger in SHR. The inhibition of PKC had no effect on constrictions in SHR, but diminished constrictions in Wistar rats. PKA inhibition slightly enhanced constrictions in Wistar rats, but reduced them in the presence of calcium in SHR. We conclude that vasoconstriction elicited by adrenergic stimulation is more dependent on extracellular calcium influx in SHR compared to Wistar rats. Moreover, the activation of PKA contributes to this calcium-dependent vasoconstriction in SHR but not in Wistar. On the other hand, PKC activation seems to play a less important role in vasoconstriction in SHR than in Wistar rats.

Protective efficacy of a filarial surface antigen in experimental filariasis.

A water-insoluble, detergent-soluble, surface-associated glycoprotein, designated as Dssd1, was found to induce microfilaria clearance in Mastomys coucha implanted with Setaria digitata. Intraperitoneal implantation of adult female worms of S. digitata in M. coucha could induce microfilaraemia lasting about 165 days in circulation. Immunization of M. coucha with Dssd1 antigen either before or after implantation of worms resulted in a significant reduction in microfilaria density. Complete clearance of circulating microfilaria was achieved by immunization (before and after implantation) in animals by 95 and 105 days post-implantation, respectively, indicating the efficacy of Dssd1 antigen in the clearance of microfilaraemia in infected M. coucha.

Spatial seroepidemiology, risk assessment and haemato-biochemical implications of bovine trypanosomiasis in low lying areas of Punjab, India.

A cross-sectional study was carried out on 594 bovines (341 buffalo adults, 31 buffalo calves, 163 cattle adults, and 59 cattle calves) to assess the exposure of native bovine population to T. evansi elicited trypanosomiasis in the low-lying areas of Punjab (India). We ruled out the endemicity of the disease with 10.77% (95%CI = 8.53-13.52) sero-positive and 23.56% (95%CI = 20.33-27.15) suspected cases by card agglutination assay. We have presented the spatial distribution of these cases as a guideline to local veterinary practitioners and policy-makers. The categorical assessment of risk factors revealed buffalo adults are the most susceptible group in the state despite insignificant differences in farm management practices. A significant increase in the WBC, platelet, AST and serum iron, and decrease in haemoglobin, haematocrit volume, and serum glucose was recorded in both T. evansi positive and suspected animals.

Seroprevalence of Neospora caninum antibodies in dogs in India.

Neospora caninum is one of most important causes of abortion in cattle worldwide, and dogs are an important risk factor for N. caninum infection in cattle. Antibodies to N. caninum were determined in 184 (126 rural, 58 urban) dogs from the Punjab State, India, using commercial monoclonal antibody-based competitive ELISA and found in 16.8% of the animals. The prevalence of N. caninum antibodies was significantly higher in rural dogs (21.4%, 27 of 126) than city dogs (6.9%, 4 of 58). To our knowledge this is the first report of N. caninum infection in canines from India.

Serological survey of antibodies to Toxoplasma gondii in sheep, cattle, and buffaloes in Punjab, India.

Sera from 186 sheep, 83 cattle, and 103 water buffaloes from Punjab, India, were evaluated for antibodies to Toxoplasma gondii using a commercial ELISA kit. This study was planned using a 2-stage random sampling procedure and sampling software 'survey toolbox.' In the first step, villages were selected randomly from a sampling frame of all the villages of Punjab, followed by selection of owners and animals in the second step. Antibodies to T. gondii were found in 7 of 186 sheep, 2 of 83 cattle, and 3 of 103 buffaloes. Results indicate a low prevalence of T. gondii in ruminants tested.

Overt bleeding in complicated P. falciparum malaria: An experience from east coast of India.

Delay in diagnosis of falciparum may result in complicated, life-threatening conditions. Though haematological abnormalities and coagulopathy are common complications that occur in malaria but complications with rare manifestations like overt bleeding do pose challenges for the clinicians worldwide. This study reports the incidence and prognosis of overt bleeding from the east coast of India and makes an attempt to relate it with the pathogenesis of the disease in severe Plasmodium falciparum malaria patients. This study was conducted in Sriram Chandra Bhanj Medical College Hospital, Cuttack, Odisha, India. A total of 120 complicated (multi organ dysfunction) malaria cases were included in this study. Amongst them 54 (45.0%) showed signs of overt bleeding and clinically the bleeding was either from one or multiple sites. Out of the total overt bleeding cases, 79.6% had elongated prothrombin time (PT), activated partial prothrombin time (aPTT) and elevated D-dimer with thrombocytopenia indicating disseminated intravascular coagulopathy (DIC). It was observed that case fatality rate was very high in coagulopathy group than the non coagulopathy group. Our observations highlight that awareness of overt bleeding in P. falcipatum infection is necessary for general practitioners in endemic areas for malaria like in Odisha for effective and timely management of complicated patients.Timely diagnosis and treatment of DIC with appropriate prescribed drugs can prevent and cure the complications of severe falciparum malaria with anti-malarial treatment.

Unraveling cryptic epizootiology of equid trypanosomosis in Punjab state of India by parasitological and sero-molecular techniques.

To unravel equid trypanosomosis caused by Trypanosoma evansi in Punjab state of India, a cross sectional study was designed by utilizing parasitological and sero-molecular tools with objective to assess the prevalence of T. evansi in association with various risk factors in all agroclimatic zones of Punjab state of India. Parasitological Romanowksy stained thin blood smears (RSTBS) to detect patent infection, molecular techniques polymerase chain reaction I (PCR I; TBR 1/2 primers; targeting minichromosomal satellite DNA of T. evansi), polymerase chain reaction II (PCR II; TR 3/4 primers; targeting variable surface glycoprotein region DNA of T. evansi) & LAMP (Loop mediated isothermal amplification) assay to detect latent infection and serological assays card agglutination test (CATT/T. evansi) & ELISA (Enzyme linked immunosorbent assay) to detect exposure status of trypanosomosis were utilized in the present study. A total 429 equid blood and serum samples from all the five agroclimatic zones of Punjab state tested by these techniques showed a prevalence of 1.39% (CL: 0-15.28) by RSTBS, 6.52% (10.94-45.09) by both TBR 1/2 PCR and LAMP assay, 5.82% (11.57-38.42) by TR 3/4 PCR, 15.15% (36.57-135.42) with CATT/T. evansi and 22.84% (17.77-840.22) with ELISA. Interpretation of various risk factors revealed that the donkey/mules population (RR = 5.46, 95% [CI] = 0.15-15.56) was found to be at higher risk of T. evansi infection predominantly at 'unorganized' farms (RR = 4.06, 95% [CI] = 0.12-4.51). Animal used for commercial purposes (RR = 3.25, 95% [CI] = 0.06-7.42), rearing of equids with other domestic animals (RR = 2.36, 95% [CI] = 0.10-17.11) and farms without application of fly repellant/insecticides/net (RR = 3.68, 95% [CI] = 0.08-5.94) made them more prone to the disease. This comprehensive report utilizing the classical, serological and molecular diagnostic tools for epidemiology of T. evansi establishes the endemic stability of this infection in all agro climatic zones of Punjab with LAMP assay to be a promisingly sensitive and specific technique for the diagnosis of T. evansi under isothermal conditions in field situations.

Prevalence of gastrointestinal parasitism in small ruminants in western zone of Punjab, India.

AIM: The aim of this study was to explore the prevalence of gastrointestinal parasitism in small ruminants in relation to various risk factors in the western zone of Punjab. MATERIALS AND METHODS: During the study, 603 fecal samples (391 of sheep and 212 of goats) were examined qualitatively by floatation and sedimentation techniques, and quantitatively by McMaster technique. RESULTS: Out of the 603 fecal (391 sheep and 212 goats) samples examined, 501 were found positive for endoparasitic infection with an overall prevalence of 83.08%, consisting of 85.16% and 79.24% in sheep and goats, respectively. Egg per gram in sheep was apparently more 1441.88±77.72 than goats 1168.57±78.31. The associated risk factors with the prevalence of gastrointestinal tract (GIT) parasites showed that females (85.97%) were significantly more susceptible than males (69.23%). Age wise the adults (>6 months) were significantly more prone to parasitic infection as compared to young ones (<6 months). Seasonal variation was recorded throughout the year and was significantly highest during monsoon (90.10%), followed by winter (83.84%) and summer (78.35%). CONCLUSION: The study revealed an overall prevalence of 83.08% of GIT parasitic infections in small ruminants constituting 85.16% in sheep and 79.24% in goats in the western zone of Punjab. The most relevant risk factors for the prevalence of gastrointestinal parasitism in ruminants were sex, age, and season.

Detection of anthelmintic resistance in sheep and goat against fenbendazole by faecal egg count reduction test.

Anthelmintic resistance against commonly used anthelmintic fenbendazole was evaluated by employing faecal egg count reduction test (FECRT) in naturally occurring gastrointestinal (GI) nematodes in the semi organized sheep and goat farms of Ludhiana and Amritsar districts. A total of 80 animals (20 each for sheep and goat in both districts) were randomly selected and their faecal samples were examined by qualitative and quantitative parasitological techniques. Results indicate presence of high level of resistance against fenbendazole in both sheep and goat population of Ludhiana and Amritsar districts. More resistance was observed in the GI nematodes from animals reared in Amritsar district as compared to Ludhiana district. The level of anthelmintic resistance observed was apparently more in sheep than goats.

Molecular survey in relation to risk factors and haemato-biochemical alteration in Theileria equi infection of equines in Punjab Province, India.

Equine piroplasmosis caused by Theileria equi, an apicomplexan intracellular haemoprotozoan parasite effects equids throughout the world. Current investigation is the first detailed epidemiological survey report based on conventional (blood smear) and primary polymerase chain reaction (PCR) for the detection of T. equi on 464 equids (426 horses and 38 donkeys/mules) in Punjab province of India. PCR targeting 18S rRNA gene of T. equi produced high fidelity 709bp amplification products with 100% concordance with blood films. The prevalence of infection was proportional (P≤0.01) to temperature and aridness of the regions, which turned out to be the most important physical factor pertaining to T. equi infection. Spatial distribution analysis revealed an increasing trend of disease prevalence from north-eastern to south-western region of Punjab by both the techniques. Haemato-biochemical analysis revealed significant decrease in total erythrocyte count and haemoglobin; and increase in mean corpuscular haemoglobin, serum globulin, glucose, creatinine, aspartate aminotransferase and bilirubin levels (P≤0.05). This study divulges the endemicity of equine theileriosis in Punjab with the prominence of various odds of infection, emphasising the pathophysiological manifestation associated with latent infection of T. equi detectable by PCR.

The Prevalence of P. malariae in Odisha, India.

Odisha, an eastern Indian state, is known to be highly endemic for malaria. The current national malaria control programme uses rapid diagnostic test (RDT) for early diagnosis and prompt treatment of the cases followed by microscopy for the identification of species. But as per the available literatures and from our own experience it is known that microscopy suffers with underestimation of mixed Plasmodium species infection, especially P. malariae and P. ovale. Since PCR has proven to have more sensitivity and specificity and able to detect multiple infection of Plasmodium species, we have made an attempt to find out the prevalence of species of parasites by PCR and compare the results with light microscopy. During this study 1589 individuals presenting with fever from two malaria endemic blocks namely Badampahar and Ghatgaon were investigated. Amongst them 110 cases were found to be PCR positive while 99 were microscopically positive for malaria. Out of the total PCR positive cases 42 cases (38.2%) had P. falciparum, 35 (31.8%) had P. vivax and 10 (9.1%) had P. malariae mono infections, while 23 (20.9 %) cases had mixed infections. Of the total 23 mixed infections 11(10%) were having P. falciparum plus P. vivax and rest 12 (10.9%) cases were having P. malariae mixed with either P. falciparum and /or P. vivax. However microscopically only 1(1.4%) case was found to have P. malariae mono infection and none of the mixed infections harbor P. malariae. Multiple kind lottery model analysis of PCR positive samples demonstrated that P. malariae is not distributed randomly in population when compared by different age groups, gender and season. The study also has shown that PCR is highly effective in screening the severe and asymptomatic malaria cases than LM. The result of this study warrants newer surveillance strategies for case detection and management by the programme.