Disinfection of environmental objects against coccidia oocysts in broiler chickens

The purpose of the research is developing a method for disinfection of environmental objects against coccidia oocysts in broiler chickens.Materials and methods. At the VNIIP – FSC VIEV vivarium, a bioassay test was performed to experimentally infect 60 chickens aged 14 days which were divided into six equivalent groups of 10 birds each and kept isolated in cages. Chickens from the first, second and third groups were administered orally, using a micropipette, 1 ml of an Eimeria oocyst suspension treated with 4, 5 and 6% solutions of the combined eimeriocide agent. Chickens from the fourth group were administered 1 ml of Eimeria oocyst suspension treated with 4% phenol solution (base drug). Chickens from the fifth group received 1 ml of a buffered solution and were used as a noninfected control. Chickens from the sixth group received 1 ml of suspension containing 2000 oocysts/mL and were used as an infected control. The efficacy of disinfection with eimeriocide and the basic drug was determined based on the percentage of decrease in the recovery of Eimeria oocysts after being exposed to drugs as compared to chickens of the infected control. The efficacy of 5% eimeriocide against poultry coccidia oocysts in a production test was determined empirically with the set of oocysts on control sites as compared with the basic drug on a poultry farm in the Moscow Region.Results and discussion. The intense-effectiveness of 4% eimeriocide against coccidia oocysts was 99.31%, and the 5 and 6% combined agent showed 100% efficacy. The basic drug, 4% phenol showed 74.65% intense-effectiveness. The results obtained in the production test of 5% eimeriocide at a dose of 0.5 l per 1 m2 with a 2 hour-exposure indicate its high efficacy for disinfection against coccidia oocysts in broiler chickens. The intense-effectiveness was 97.25% versus 59.03% efficacy of the base drug, phenol.

Anticoccidial Activity of Qinghao Powder Against Eimeria tenella in Broiler Chickens

Artemisia annua (AAH) is traditionally used as an anti-malarial, expectorant and antipyretic Chinese medicine. The aim of this study was to explore the therapeutic effect of Qinghao Powder (QHP) on chicken coccidiosis, evaluate the safe dosage of QHP, and provide test basis for clinical medication. High-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) were used to detect artemisinin in Qinghao Powder (QHP) for quality control. The level of artemisinin in QHP was 81.03 mg/g. A total of 210 chicks (14 days of age) were divided randomly into seven groups: three QHP treatments (0.15, 0.30, and 0.60 g/kg), a toltrazuril control (1.00 mL/L), a sulfachloropyrazine sodium control (SSC, 0.30 g/L), an E. tenella-infected control, and a healthy control group. All the groups were inoculated orally with 7 × 104E. tenella oocysts except for the healthy control group. After seven days of administration, compared with the infected control group, chicks which were administered QHP, SS, and toltrazuril showed less bloody feces, oocyst output, and cecal lesions, and the protection rates were improved. The maximum rBWG and ACI were found in the SS-medicated group, followed by the groups medicated with 0.60 and 0.30 g/kg QHP. Therefore, a 0.30 g/kg dose level of QHP in the feed was selected as the recommend dose (RD) in the target animal safety test, in which 80 broiler chicks (14 days of age) were randomly divided into four major groups (I-healthy control group; II-1× RD; III-3× RD; IV-6× RD), with each group subdivided into two subgroups (A and B) consisting of 10 chicks each. After 7-day (for sub-group A) or 14-day (for sub-group B) administration, compared with the healthy control, treatment-related changes in BWG, feed conversion ratio (FCR), relative organ weight (ROW) of the liver, WBC counts, and levels of RBC, HGB, ALT, AST, and TBIL were detected in the 3× and 6× RD groups. No differences were noted in necropsy for all doses, and histopathological examinations exhibited no QHP-associated signs of toxicity or abnormalities in the liver or kidney. The findings suggest that QHP at a dose of 0.30 g/kg feed would be appropriate for therapy and intermittent treatment of E. tenella-infected chicks, the dosage in clinical applications should be set according to the recommended dose to ensure animal safety.

Type I Diabetes Mellitus Increases the Cardiovascular Complications of Influenza Virus Infection

People with diabetes mellitus are susceptible to both cardiovascular disease and severe influenza A virus infection. We hypothesized that diabetes also increases risks of influenza-associated cardiac complications. A murine type 1 (streptozotocin-induced) diabetes model was employed to investigate influenza-induced cardiac distress. Lung histopathology and viral titres revealed no difference in respiratory severity between infected control and diabetic mice. However, compared with infected control mice, infected diabetic mice had increased serum cardiac troponin I and creatine-kinase MB, left ventricular structural changes and right ventricular functional alterations, providing the first experimental evidence of type I diabetes increasing risks of influenza-induced cardiovascular complications.

MOSQUITO-BORNE PARASITES IN PATIENTS NEWLY INFECTED WITH HIV IN RELATIONSHIP WITH CD4 COUNT AND TNF ΑLPHA

Background: Mosquito-borne parasites include the pathogenic protozoa and helminths that are transmitted by the insect vector which may co-infect with other organisms to elicit an immune response. Purpose: To determine the frequency of mosquito-borne parasites in patients newly infected with HIV in relationship with CD4 count and TNFα. Method: Thirty-one (31; aged 15-32 years; male-12; female-19) newly diagnosed HIV positive patients and fifty (50) age-matched HIV negative volunteers were recruited as a control subject for this study. All subjects were negative to anti-HCV/HBsAg ELISA, Plasmodium, Acid-Fast Bacilli (AFB) tests and the control subjects were also negative to HIVP24 Ag-Ab ELISA, Plasmodium spp. and Wuchereria bancrofti microscopy. Venous blood including Night blood samples and sputum samples were obtained from the participants for CD4 count by cyflowmetry, TNFα, HIVP24Ag-Ab, anti-HCV, HBsAg by ELISA and microscopic identification by Giemsa staining while Sputum sample was used for Ziehl Neelsen staining to demonstrate Acid Fast Bacilli (AFB). Result: A lower frequency of 25.8% (Rajan, 2008) Plasmodium spp. and 6.5% (James et al., 2015) W. bancrofti was obtained in newly infected HIV patients compared with 32% (Zeitlmann et al., 2001) Plasmodium spp. and 8% (WHO, 2019) W. bancrofti obtained in the non-HIV infected control subjects. Showed a significant decrease in CD4 count and increase in plasma TNFα in both HIV mono-infection and coinfection with Plasmodium spp. and W. bancrofti compared with the results obtained in the non-HIV infected control subjects (p<0.05) and the results obtained in the newly infected HIV patients without Plasmodium spp. and W. coinfection (p<0.05). Conclusion: There was a significant increase in plasma TNFα and a decrease in CD4 count in both HIV mono-infection and coinfection with Plasmodium spp. and W. bancrofti while a lower frequency of Plasmodium spp. and W. bancrofti was obtained in newly infected HIV patients compared with the results obtained in the non-HIV infected control subjects.

EFFICIENCY OF THE COMPLEX DRUG CYSTODEZ-ULTRA AGAINST EIMERIA SPP. OOCYSTS IN BROILER CHICKENS

The practice of successful poultry farms shows that the prevention of poultry parasitoses includes a complex of therapeutic and prophylactic measures aimed both against exogenous oocysts in the environment and against endogenous stages of the pathogen inside the bird's body with the use of modern antiparasitic drugs. In view of the foregoing and special resistance of Eimeria oocysts in the environment, it is possible to create an effective disinfectant against them using several active ingredients and excipients. Among these drugs, we should note glutaraldehyde, crystalline iodine and potassium iodide when used together at optimal concentrations. The experiments were carried out from August to October 2019 in the laboratory and vivarium of the Institute on 60 14-day-old Cobb-500 broiler chickens. Working solutions of Cystodez-ultra in different concentrations were prepared in advance, as well as a buffer solution WSH for preserving bird feces. Cystodez-ultra is a complex drug in a liquid form for disinfection of veterinary supervision objects and prevention of invasive diseases of birds. In the test, the chickens of the first, second and third groups were administered 1 ml of a suspension of Eimeria spp. oocysts treated with 3, 4 and 5% solutions of Cystodez-ultra orally using a micropipette; the fourth group was also administered 1 ml of a suspension of Eimeria oocysts treated with Phenol solution 4%. Broilers of the fifth group were administered a suspension with sporulated Eimeria spp. oocysts at a dose of 2000 per 1 ml not treated with a disinfectant - the infected control, and the sixth group 1 ml of WSH buffer solution - non-infected control. During the experiment, the chickens were followed up daily paying attention to their general condition, behavior, appetite, and visible physiological changes. According to the study results, the intense-effectiveness of Cystodez-ultra in 3, 4 and 5% concentration against sporulated Eimeria oocysts was 93.08, 100 and 100%, respectively, and Phenol in 4% concentration 76.92%. The number of oocysts in 1g of feces in the infected control was 4680 specimens, and the chickens of the uninfected control remained free from the invasion at all times of the study.

The Roles of Betulinic Acid on Circulating Concentrations of Creatine Kinase and Immunomodulation in Mice Infected With Chloroquine-Susceptible and Resistant Strains of Plasmodium Berghei

Complete malarial therapy depends largely on the immunological and inflammatory response of the host to the invading potentials of malarial parasite. In this study, we evaluated the roles of betulinic acid on immunological response, anti-inflammatory potentials and concentrations of creatine kinase in mice infected with chloroquine susceptible (NK 65) and resistant (ANKA) strains of Plasmodium berghei. Serum Interlukins 1β and 6 (IL-1 β, IL-6), tumour necrosis factor alpha (TNFα), immunoglobulins G and M (IgG and IgM), C-reactive protein (CRP) and creatine kinas (CK). Furthermore, liver marker enzymes; aspartate, alanine aminotransferases (AST and ALT, respectively) and gamma glutammyl transferase (GGT) were determined. The results showed that betulinic acid dose dependently decreased IL-1 β, IL-6, TNFα and CRP relative to the infected control. The IgG and IgM levels significantly increased in both models while CK decreased insignificantly in both models. Serum AST, ALT and GGT significantly decreased compared to the infected control. These results showed that betulinic acid has anti-inflammatory, immunomodulatory and mitigating effects on malarial infection in mice. Furthermore, the down-regulatory effect of betulinic acid on CK is indicative of decrease in muscle injury which is a major pathological concern in malarial infection and treatment.

The Curative Effect of Zingiber Officinale and Cinnamomum Zeylanicum Extracts on Experimental Trichinella Spiralis Infection

Trichinellosis is a re-emerging zoonotic disease that has become a public health concern since its reported human outbreaks in many countries. The traditional therapy has many adverse effects in addition to the developing resistance. So, this necessitates finding effective natural alternatives. The current study targeted to assess the potential therapeutic effects of Zingiber officinale and Cinnamomum zeylanicum in comparison to albendazole, a conventional therapy for treatment of trichinosis. Sixty mice were classified into five groups (12 mice each), non-infected control, infected control, combined albendazole and prednisolone, Zingiber officinale, and Cinnamomum zeylanicum treated groups. Mice sacrifice was performed on the 7th and 35th days post infection for intestinal and muscular phases respectively. Efficiency of the used preparations was assessed by parasitological, histopathological, immunohistochemical, biochemical studies in addition to ultrastructural evaluation using transmission electron microscopy. A significant reduction in the mean number of T. spiralis adults and larvae was observed in Zingiber officinale and Cinnamomum zeylanicum treated groups, (64.5%, 50.8%) and (68%, 54.6%) respectively. Also, both extracts showed moderate cytoplasmic reactivity for TGF-β1, (69.3% & 67.8%) respectively. The highest reduction in serum TNF- α level was observed in Zingiber officinale treated group during the muscle phase (58.4%) while in the intestinal phase was 50%. The ultrastructural study revealed degenerative effects on both adults and larvae in addition to obvious improvement of the histopathological changes in the small intestine and muscles. We concluded that these herbal extracts especially Zingiber officinale can be considered a practical and successful alternative for the treatment of trichinellosis.

Efficacy of a multivalent vaccine against Fasciola hepatica infection in sheep

In this work we report the protection found in a vaccination trial performed in sheep with two different vaccines composed each one by a cocktail of antigens (rCL1, rPrx, rHDM and rLAP) formulated in two different adjuvants (Montanide ISA 61 VG (G1) and Alhydrogel®(G2)). The parameters of protection tested were fluke burden, faecal egg count and evaluation of hepatic lesions. In vaccinated group 1 we found a significant decrease in fluke burden in comparison to both unimmunised and infected control group (37.2%; p = 0.002) and to vaccinated group 2 (Alhydrogel®) (27.08%; p = 0.016). The lower fluke burden found in G1 was accompanied by a decrease in egg output of 28.71% in comparison with the infected control group. Additionally, gross hepatic lesions found in vaccine 1 group showed a significant decrease (p = 0.03) in comparison with unimmunised-infected group. The serological study showed the highest level for both IgG1 and IgG2 in animals from group 1. All these data support the hypothesis of protection found in vaccine 1 group.

Diclazuril-induced expression of CDK-related kinase 2 in the second-generation merozoites of Eimeria tenella

Background: Diclazuril is a classic anticoccidial drug. The key molecules of diclazuril in anticoccidial action allows target screening for the development of anticoccidial drugs. In the present study, a diclazuril anticoccidiosis animal model was established, and the transcription and translation levels of the CDK-related kinase 2 of Eimeria tenella (EtCRK2) were detected through quantitative real-time PCR and Western blot analysis, respectively. The localisation of EtCRK2 in merozoites was examined with immunofluorescence techniques.Results: The mRNA and protein expression levels of EtCRK2 decreased in the infected/diclazuril group compared with those in the infected/control group. In addition, immunofluorescence analysis showed that EtCRK2 was localised in the cytoplasm of the merozoites. The fluorescence intensity of EtCRK2 in the infected/diclazuril group was significantly weaker than that in the infected/control group.Conclusions: The anticoccidial drug diclazuril against E.tenella affects the expression pattern of EtCRK2 molecule, and EtCRK2 is a potential target for new drug development.

Diclazuril-Induced Expression of CDK-Related Kinase 2 in the Second-Generation Merozoites of Eimeria Tenella

Background：Coccidiosis caused by Eimeria tenella infection, directly or indirectly leads to great loss to poultry industry. With the emergence of drug-resistance in chicken coccidia, it is imperative to develop new drugs. Cyclin-dependent kinases (CDKs) regulate cell cycle progression in numerous organisms by acting as key molecular switches. Results: In the present study, a diclazuril anticoccidiosis animal model was established and CDK-related kinase 2 (CRK2) in the second-generation merozoite of E. tenella (EtCRK2) gene was amplified through reverse transcription-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli Rosetta (DE3). Purified recombinant protein was used for antiserum preparation. Subsequently, EtCRK2 transcription and translation levels were detected through quantitative real-time PCR and Western blot analysis, respectively. The localization of EtCRK2 in merozoites was examined via immunofluorescence techniques. Results showed that the mRNA and protein expression levels of EtCRK2 decreased in the infected/diclazuril group compared with those in the infected/control group. In addition, immunofluorescence analysis showed that EtCRK2 was localized in the cytoplasm of merozoites. The fluorescence intensity of EtCRK2 in the infected/diclazuril group was significantly weaker than that in the infected/control group. Conclusions: This study demonstrated that the anticoccidial drug diclazuril against E. tenella by affecting the expression pattern of EtCRK2 molecule, and EtCRK2 may be used as a candidate target for new drug development.