Use of FAMACHA to Detect Anaemia and Control of Gastrointestinal Parasite in Goats of A and N Islands, India

Background: FAffaMAlanCHArt (FAMACHA) is a technique for treating the animals infested with gastrointestinal parasites by using clinical anaemia as the determinant. The test is based on the assessment of colour variation of the conjunctival mucous membrane. Methods: In the present study FAMACHA has been standardized and used to correlate the anaemic status of goats with faecal egg count (gastro intestinal parasites) and blood profile. On the basis of FAMACHA score and faecal egg count, the goats were treated with benzimidazole compound and macrocyclic lactone. Result: The result indicated significant (P less than 0.05) reduction in the faecal egg count of Haemonchus spp from 1200±155.76 to 525±75, Strongyloides spp from 469.57±77.37 to 78.26±33.83 and Eimeria spp from 900±240.05 to 195.65±55.34 respectively. FAMACHA score of anaemic goats improved from 4.08±0.11 to 3.2±0.13. Mean haemoglobin (g/dl), packed cell volume (PCV), red blood cell (RBC) and white blood cell (WBC) improved from 8.33 to 8.99, 22.66 to 24.86, 13.73 to 14.76 and 18029 to 19894, respectively which indicated clinical improvement. Highly significant (P less than 0.01) correlation was observed between the FAMACHA score and the haemoglobin level. It could be concluded that FAMACHA is a very handy tool to detect anaemic status of the animals and correlate with gastrointestinal parasitic load.

​Faecal Consistency: A Measure of Gastrointestinal Parasitic Infections in Goats

Background: Gastrointestinal (GI) parasitism in goats is a serious and widely distributed problem. Diarrhoea, moderate or severe, though a result of various factors involving feed, fodder, seasons and state of gut physiology of digestion, is a common symptom in goats infested with GI parasites, the nematodes and coccidia. Intensity of diarrhoea corresponds to quantum and type of parasitism. Methods: In the present study, 1451 faecal samples collected from goats of different ages ( less than 3 M, greater than 3-6 M, greater than 6-12 M and greater than 12 M) were categorized as normal, lumpy and diarrheic based on their consistency and were processed for parasitic infections. The GI parasitic load was quantified by faecal eggs/oocysts counting (FEC and FOC) per gram of faeces through the modified Mac-master technique. Identification of parasites was based on the morphology of third-stage larvae procured by coproculture of randomly selected positive samples. FEC/FOC data (strongyle and coccidian), being skewed and uneven, were log-transformed to normalize. The transformed data [loge (100+n)] was analyzed statistically and results were interpreted. Result: Means of FEC and FOC in three type of faecal samples viz. normal, lumpy and diarrheic, were significantly different. Similarly, the effect of age on FEC and FOC was significant. The interaction between animals age × faecal sample type was significant in coccidian and non-significant in strongyle infection. The results were discussed to support the hypothesis that faecal consistency was a measure of coccidian and strongyle worm load in goats and can be a basis of targeted selective treatment.

Eurytrema coelomaticum infection: correlation between parasite burden and impairment of pancreatic exocrine enzyme secretion

ABSTRACT: Eurytrema coelomaticum is a trematode reported in the pancreatic ducts of ruminants. It is conjectured that may cause disorders in the pancreas, as well as digestive and metabolic processes dependent on them. This study, determined if there is an impairment of exocrine pancreatic function, and correlated it with parasite burden. Pancreas, blood, and fecal samples were collected from 119 bovines at a abattoir. Stool samples were subjected to the gelatin and x-ray film digestion tests (to detect the presence of trypsin in feces). Using blood samples, the following biochemical tests were performed: amylase, lipase, glucose, fructosamine, cholesterol, triglycerides, total protein, albumin, and globulins. Analyses were correlated with pancreatic parasite burden. Cattle with a high parasitic load presented higher incidence of negative tests in both gelatin digestion and x-ray film digestion tests (P < 0.001) when compared to non-parasitized animals and those with a low parasitic load. Changes in those tests only occurred if the parasitemia was moderate or severe. The activity of the amylase and lipase enzymes was significantly higher in animals with low parasitemia (P < 0.05), compared to non-parasitized animals and with a high parasitic burden. In this study, in cases of high parasitemia, negative results were observed in both gelatin and x-ray film in the feces digestion tests. However, the low infection of E. coelomaticum, higher levels of serum amylase and lipase that also indicated loss of pancreatic exocrine functions were reported.

Long-Term In Vitro Passaging Had a Negligible Effect on Extracellular Vesicles Released by Leishmania amazonensis and Induced Protective Immune Response in BALB/c Mice

Depending on Leishmania species and the presence/absence of virulence factors, Leishmania extracellular vesicles (EVs) can differently stimulate host immune cells. This work is aimed at characterizing and evaluating the protective role of EVs released by Leishmania amazonensis promastigotes under different maintenance conditions. Initially, using a control strain, we standardized 26°C as the best release temperature to obtain EVs with a potential protective role in the experimental leishmaniasis model. Then, long-term (LT-P) promastigotes of L. amazonensis were obtained after long-term in vitro culture (100 in vitro passages). In vivo-derived (IVD-P) promastigotes of L. amazonensis were selected after 3 consecutive experimental infections in BALB/c mice. Those strains developed similar lesion sizes except for IVD-P at 8 weeks post infection. No differences in EV production were detected in both strains. However, the presence of LPG between LT-P and IVD-P EVs was different. Groups of mice immunized with EVs emulsified in the adjuvant and challenged with IVD-P parasites showed decreased lesion size and parasitic load compared with the nonimmunized groups. The immunization regimen with two doses showed high IFN-γ and IgG2a titers in challenged mice with either IVD-P or LT-P EVs. IL-4 and IL-10 were detected in immunized mice, suggesting a mixed Th1/Th2 profile. EVs released by either IVD-P or LT-P induced a partial protective effect in an immunization model. Thus, our results uncover a potential protective role of EVs from L. amazonensis for cutaneous leishmaniasis. Moreover, long-term maintenance under in vitro conditions did not seem to affect EV release and their immunization properties in mice.

Evaluation of Trypanosoma cruzi parasitic load by real-time PCR and blood culture in long-term kidney transplant recipients

Introduction: Acute Chagas disease involving reactivation can occur after organ transplant, and follow-up by direct parasitological or molecular methods is essential for monitoring the parasitic load in such patients. In contrast, there is a little data on the parasitic load in long-term organ recipients. In this study, we examined the parasitic load in long-term kidney transplant patients and assessed the possibility of late Chagas disease reactivation. Methodology: Blood cultures and real-time PCR were used to assess the parasitic load in four immunosuppressed patients who underwent kidney transplants (between 1996 and 2014) and were also treated for parasites. Results: There were no positive blood culture or real-time PCR results in Chagas disease patients who received kidney transplants. The real-time PCR presented detection limit of 0.1 parasite equivalent/mL. The time interval between the transplant and sample collection varied from one to 19 years. Conclusions: No parasites were detected in the evaluated patients. The use of benznidazole and immunosuppressive therapy may have contributed to control the T. cruzi infection. In transplanted patients with Chagas disease, the use of methods such real-time PCR and blood culture can monitor the parasitic load and prevent disease reactivation.

Investigating in Vivo and in Vitro Effects of Ethanolic and Aqueous Extracts of Myrtle (Myrtus communis) on Leishmania major

Background: The extract of myrtle plant contains polyphenolic compounds that show antibacterial, antiviral, and anti-parasitic properties. We aimed to investigate the therapeutic effect of aqueous and ethanolic myrtle extract against leishmaniasis caused by L. major in vivo and in vitro conditions. Methods: This study was carried out in Tarbiat Modares University, Tehran, Iran in 2018. Aqueous and ethanolic extract of myrtle plant at 6.25 to 400 mg/ml concentrations were tested on Leishmania major promastigotes, non-infected macrophages, and macrophages infected with amastigotes in vitro using counting, MTT and flow cytometry techniques. Then, BALB/c mice were treated with ethanolic, aqueous and a mixture of both extracts of myrtle plant. The treatment was carried out for four weeks. Then, the effectiveness of the herbal medicine was assessed by measuring wounds diameters, mice weights and their mortality rate on weekly basis. Results: The IC50 values of aqueous and ethanolic extracts for promastigotes were 7.86 and 11.66 μg/mL respectively. The IC50 values of the aqueous and ethanolic extracts for amastigotes were 12.5 and 47.2 μg/mL respectively. Flow cytometry indicates 62.88% and 60.16% apoptosis induced by ethanolic and aqueous extract of myrtle plant respectively. The lowest parasitic load was seen in the group treated with ethanolic extract. Conclusion: The lesion sizes for treated groups with extracts were similar to those treated with glucantime. Oral administration instead of injection is another advantage of myrtle plant over glucantime, which makes the herb easy and more practical.

Study on incidence and pathology of gastrointestinal parasitic infections in Nilgai Boselaphus tragocamelus in Hisar, Haryana, India

The present study was conducted on 20 dead carcasses of Nilgai Boselaphus tragocamelus brought to the Department of Veterinary Pathology, Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS), Hisar for post mortem examination. Thorough necropsy examinations were conducted and the representative samples for parasitic examination were collected. Most of the carcasses showed varying degrees of traumatic injuries (9), external wounds and haemorrhages (5) along with putrefactive changes (6). Intestinal contents and faecal samples were collected and screened for the presence of any parasite by sedimentation and flotation techniques. Out of 20 faecal samples, overall incidence of parasitic gastrointestinal tract infection was 40% (8/20). Out of 20, eight cases revealed presence of coccidian oocysts, however, among the eight cases, one case also revealed mixed infection of Moneizia andStrongylespp.,andanother case with Strongyle and Trichuris spp. Gross pathology of the intestines revealed varying degrees of vascular changes like petechial haemorrhages and the presence of catarrhal exudate. Histopathological examination revealed mild to moderate congestion, fused villi, desquamated mucosal epithelium in focal areas, and infiltration of mononuclear cells mainly lymphocytes. The different developmental stages of coccidian spp.were also observed in the intestinal mucosa. In conclusion, the preliminary study reported the parasitic load and pathological changes in the intestinal tract which further indicated the parasitism of these wild animals which is of immense significance from the epidemiological point of view.

Insights into the Relation between Oxidative Stress and Malaria: A Mechanistic and Therapeutic Approach

The mortality rate due to malaria has increased tremendously in the last decade. Even though the causative agent of this disease is known, the preventive measures are not potent enough to control the spread of this disease. Malarial infection involves a strong interrelationship between oxidative stress and pathogenesis. This review addresses the various oxidative stress-related mechanisms associated with vector defense, host immunity, plasmodial pathogenesis, and corresponding therapeutic strategies. The mechanisms involving host and vector defense show both similarity and contradiction to the processes involving plasmodial pathogenesis under different circumstances. Therefore, corresponding ameliorative peculiarities are observed in the therapeutic mechanisms adopted by the anti-malarial drugs. The malarial parasite augments oxidative stress to weaken the host and exerts antioxidant effects against host defense mechanisms. However, the anti-malarial drugs induce oxidative insult to reduce parasitic load and exert antioxidant effects against parasite infection-induced oxidative stress in host. Thus, the anti-malarial drugs exhibit antioxidant activity in hosts and/or pro-oxidant activity in parasites.

The Crimean-Congo haemorrhagic fever tick vector Hyalomma marginatum in south of France: Modelling its distribution and determination of factors influencing its establishment in a newly invaded area

For the first time we built a correlative model for predicting the distribution of H. marginatum, one of the main vector of Crimean-Congo Haemorrhagic fever virus (CCHFV), at high resolution in a recently colonized area, namely south of France. Field tick collections were conducted on horses from 2016 to 2021 in 14 French southern departments, which resulted in a first map of H. marginatum on the national territory. Such updated presence/absence data, as well as the mean number of H. marginatum per examined animal (mean parasitic load) as a proxy of the tick abundance, were correlated to multiple parameters that described the climate and habitats characterizing each collection site, as well as movements of horses as a possible source of tick diffusion and new establishment. Our model highlighted the importance of warm temperatures all along the year, as well as dry conditions during summer and moderate annual humidity for the establishment of H. marginatum. A predominance of open natural habitats in the environment was also identified as a supporting factor, in opposition to artificial and humid habitats that were determined as unsuitable. Based on this model, we predicted the current suitable areas for the establishment of the tick H. marginatum in South of France, with a relatively good accuracy using internal and external validation methods. Concerning tick abundance, some correlative relationships were similar than in the occurrence model but the type of horse movements were also pointed out as an important factor explaining the mean parasitic load, leading to differential exposure to ticks. The limitations of estimating and modelling H. marginatum abundance in a correlative model are discussed.

Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification of Leishmania Parasites Causing Most Leishmaniasis in Iran

Many laboratory methods are used to diagnose leishmaniasis because it is characterized by varied symptoms and caused by different Leishmania species. A quantitative real-time PCR method based on a TaqMan probe was developed and modified for accurate identification of human cutaneous leishmaniasis (caused by Leishmania major or Leishmania tropica) from endemic areas of Iran. Two gene regions of amino acid permease 3 (AAP3) and cytochrome oxidase II (COII) were considered. Six new sets of species-specific primers and probes were designed. A total of 123 samples were examined and employed to evaluate and validate real-time PCR. According to parasitic load of the genesig®Leishmania Advanced Standard Kit, a serial dilution of purified plasmid (2–2×107 copies/reaction) was prepared under the same conditions for both genes. Specific primers and probes were able to detect three and six parasite copies in AAP3 and COII genes, respectively, and were able to detect three copies of parasites for L. major and L. tropica. The sensitivities of the reference kit and our method were 98.7 and 98.1%, respectively, and specificity was 100% for detecting parasite genomes in all assays. Designed primers and probes performed well in terms of efficiency and regression coefficient. For AAP3 and COII genes, respectively, the linear log range was 7 and the correlation coefficient (R2) was 0.749 and 0.996 for the reference kit using the standard generated curve and 0.98 and 0.96 with serial dilutions of parasite DNA. This research detected L. major and L. tropica definitely and opens the horizon for the other scientists in the multiplex reactions in designing and optimization of the conditions in silico and in vivo.