Objective: This research aims to study the target specificity of selective bioactive compounds in complexing with the human angiotensin-converting enzyme (hACE2) receptor to impede the severe acute respiratory syndrome coronavirus 2 influx mechanism resulting in cardiac injury and depending on the receptor’s active site properties and quantum tunneling.  Materials and Methods: A library of 120 phytochemical ligands was prepared, from which 5 were selected considering their absorption, distribution, metabolism, and excretion (ADMET) and quantitative structure–activity relationship (QSAR) profiles. The protein active sites and belong­ing quantum tunnels were defined to conduct supramolecular docking of the aforementioned ligands. The hydrogen bond formation and hydrophobic interactions between the ligand–recep­tor complexes were studied following the molecular docking steps. A comprehensive molecular dynamic simulation (MDS) was conducted for each of the ligand–receptor complexes to figure out the values – root mean square deviation (RMSD) (Å), root mean square fluctuation (RMSF) (Å), H-bonds, Cα, solvent accessible surface area (SASA) (Å2), molecular surface area (MolSA) (Å2), Rg (nm), and polar surface area (PSA) (Å). Finally, computational programming and algorithms were used to interpret the dynamic simulation outputs into their graphical quantitative forms.  Results: ADMET and QSAR profiles revealed that the most active candidates from the library to be used were apigenin, isovitexin, piperolactam A, and quercetin as test ligands, whereas serpentine as the control. Based on the binding affinities of supramolecular docking and the parameters of molecular dynamic simulation, the strength of the test ligands can be classified as isovitexin > quercetin > piperolactam A > apigenin when complexed with the hACE2 receptor. Surprisingly, serpentine showed lower affinity (−8.6 kcal/mol) than that of isovitexin (−9.9 kcal/mol) and quer­cetin (−8.9 kcal/mol). The MDS analysis revealed all ligands except isovitexin having a value lower than 2.5 Ǻ. All the test ligands exhibited acceptable fluctuation ranges of RMSD (Å), RMSF (Å), H-bonds, Cα, SASA (Å2), MolSA (Å2), Rg (nm), and PSA (Å) values.  Conclusion: Considering each of the parameters of molecular optimization, docking, and dynamic simulation interventions, all of the test ligands can be suggested as potential targeted drugs in blocking the hACE2 receptor. J. Adv. Vet. Anim. Res., 8(4): 540-556, December 2021 http://doi.org/10.5455/javar.2021.h544

Objective: This research aims to study the target specificity of selective bioactive compounds in complexing with the human angiotensin-converting enzyme (hACE2) receptor to impede the severe acute respiratory syndrome coronavirus 2 influx mechanism resulting in cardiac injury and depending on the receptors active site properties and quantum tunneling. Materials and Methods: A library of 120 phytochemical ligands was prepared, from which 5 were selected considering their absorption, distribution, metabolism, and excretion (ADMET) and quantitative structureactivity relationship (QSAR) profiles. The protein active sites and belong¬ing quantum tunnels were defined to conduct supramolecular docking of the aforementioned ligands. The hydrogen bond formation and hydrophobic interactions between the ligandrecep¬tor complexes were studied following the molecular docking steps. A comprehensive molecular dynamic simulation (MDS) was conducted for each of the ligandreceptor complexes to figure out the values root mean square deviation (RMSD) (Å), root mean square fluctuation (RMSF) (Å), H-bonds, Cα, solvent accessible surface area (SASA) (Å2), molecular surface area (MolSA) (Å2), Rg (nm), and polar surface area (PSA) (Å). Finally, computational programming and algorithms were used to interpret the dynamic simulation outputs into their graphical quantitative forms. Results: ADMET and QSAR profiles revealed that the most active candidates from the library to be used were apigenin, isovitexin, piperolactam A, and quercetin as test ligands, whereas serpentine as the control. Based on the binding affinities of supramolecular docking and the parameters of molecular dynamic simulation, the strength of the test ligands can be classified as isovitexin > quercetin > piperolactam A > apigenin when complexed with the hACE2 receptor. Surprisingly, serpentine showed lower affinity (−8.6 kcal/mol) than that of isovitexin (−9.9 kcal/mol) and quer¬cetin (−8.9 kcal/mol). The MDS analysis revealed all ligands except isovitexin having a value lower than 2.5 Ǻ. All the test ligands exhibited acceptable fluctuation ranges of RMSD (Å), RMSF (Å), H-bonds, Cα, SASA (Å2), MolSA (Å2), Rg (nm), and PSA (Å) values. Conclusion: Considering each of the parameters of molecular optimization, docking, and dynamic simulation interventions, all of the test ligands can be suggested as potential targeted drugs in blocking the hACE2 receptor. [J Adv Vet Anim Res 2021; 8(4.000): 540-556]

Objective: The present study aimed to explain the effect of fetal bovine serum (FBS) on the in vitro production of porcine embryos and the molecular effects of FBS on the growing of porcine embryos.  Materials and Methods: Immature porcine oocytes were matured and fertilized in vitro. The resulting zygotes were cultured in porcine zygotic medium-3- until day 7 and FBS was added on day 4. Without FBS, it was treated as a control group. Quantitative real-time PCR and 2′,7′-dichloro-di­hydro-fluorescein diacetate (H2DCFDA) molecular staining techniques were used to detect the expression patterns of apoptosis-associated genes and the accumulation of reactive oxygen spe­cies (ROS), respectively. Paired student’s t-test was used by GraphPad Prism statistical software.  Results: FBS supplementation boosted blastocyst (BL) development and total cell count per BL substantially (p < 0.05). However, hatching and hatched BLs also increased in the FBS-treated group compared to the control. We also found that ROS accumulation in FBS-treated embryos was significantly reduced (p < 0.05) compared to the control group. The expression of the anti-apop­totic gene BCL-2 was significantly increased in FBS-treated BLs, but the pro-apoptotic gene, caspase-3 expression, was significantly reduced in FBS-treated BLs.  Conclusion: Our results suggest that FBS supplementation in porcine culture media could increase porcine embryo production by decreasing ROS accumulation and increasing the anti-apoptotic gene expression in developing BLs. J. Adv. Vet. Anim. Res., 8(4): 589-596, December 2021 http://doi.org/10.5455/javar.2021.h549

Objective: The present study aimed to explain the effect of fetal bovine serum (FBS) on the in vitro production of porcine embryos and the molecular effects of FBS on the growing of porcine embryos. Materials and Methods: Immature porcine oocytes were matured and fertilized in vitro. The resulting zygotes were cultured in porcine zygotic medium-3- until day 7 and FBS was added on day 4. Without FBS, it was treated as a control group. Quantitative real-time PCR and 2′,7′-dichloro-di¬hydro-fluorescein diacetate (H2DCFDA) molecular staining techniques were used to detect the expression patterns of apoptosis-associated genes and the accumulation of reactive oxygen species (ROS), respectively. Paired students t-test was used by GraphPad Prism statistical software. Results: FBS supplementation boosted blastocyst (BL) development and total cell count per BL substantially (p < 0.05). However, hatching and hatched BLs also increased in the FBS-treated group compared to the control. We also found that ROS accumulation in FBS-treated embryos was significantly reduced (p < 0.05) compared to the control group. The expression of the anti-apoptotic gene BCL-2 was significantly increased in FBS-treated BLs, but the pro-apoptotic gene, caspase-3 expression, was significantly reduced in FBS-treated BLs. Conclusion: Our results suggest that FBS supplementation in porcine culture media could increase porcine embryo production by decreasing ROS accumulation and increasing the anti-apoptotic gene expression in developing BLs. [J Adv Vet Anim Res 2021; 8(4.000): 589-596]

Objective: This study was conducted to generate data on the prevalence, species composition, and factors associated with small ruminant nematode infection in the South Omo zone, in South-western Ethiopia.  Material and Methods: A cross-sectional study through fecal nematode parasite examination was conducted. Simple floatation test and coproculture, followed by Baermann technique, were used for nematode egg and third-stage larvae (L3) separation and identification. The McMaster method was used to calculate the egg per gram of feces (EPG).  Results: From a total of 242 sheep and goats examined, 72.34% were infested with single or mixed nematode parasites. District, “Kebele”, species, body condition score, and age were significantly (p < 0.05) associated with nematode infestation. Simple logistic regression analysis indicated that nematode infestation in Bena-Tsemay district (78.33%) was significantly (p < 0.05) higher by a factor of 0.54 (OR 95% CI: 0.30–0.96) than Hamer district (66.39%). Among the species, caprine (79.43%) was significantly (p < 0.05) infested than ovine (62.37%) by a factor of 0.45 (OR 95% CI: 0.25–0.81). Moreover, infestation on poor (79.12%) and medium (70.96%) body conditioned animals was higher by a factor of 2.94 (OR 95% CI: 1.41–6.26) and 1.76 (OR 95% CI: 0.88–3.53) than on good body conditioned animals (63.79%). Likewise, infestation in age groups of 1–3 years (78.66%) and >3 years (84.40%) was significantly (p < 0.05) higher by a factor of 4.83 (OR 95% CI: 2.31–10.46) and 8.23 (OR 95% CI: 3.98–17.75) than younger age groups (41.37%), respectively. A moderate parasitic burden was observed on 52.90% of gastrointestinal nematodes (GIN)-infested animals with significantly (p < 0.05) higher EPG in females than males. Furthermore, mixed infes­tation of Trichostrongylus axei and Eimeria (6.19%), Haemonchus contortus and Eimeria (5.78%), and Trichostrongylus vitrines and Eimeria (5.78%) were dominantly identified. On the contrary, T. axei (15.70%), Eimeria (8.67%), H. contortus (7.43%), and Trichostrongylus colubriformis (7.02%) were dominant single infestations.  Conclusion: The current study revealed the highest prevalence of GIN in the study area, which needs strategic control, needs to enhance community awareness toward GIN control and pre­vention, and to implement further investigation into anthelminthic efficacy to solve the problem. J. Adv. Vet. Anim. Res., 8(4): 597-605, December 2021 http://doi.org/10.5455/javar.2021.h550

Objective: This study was conducted to generate data on the prevalence, species composition, and factors associated with small ruminant nematode infection in the South Omo zone, in South-western Ethiopia. Material and Methods: A cross-sectional study through fecal nematode parasite examination was conducted. Simple floatation test and coproculture, followed by Baermann technique, were used for nematode egg and third-stage larvae (L3) separation and identification. The McMaster method was used to calculate the egg per gram of feces (EPG). Results: From a total of 242 sheep and goats examined, 72.34% were infested with single or mixed nematode parasites. District, Kebele, species, body condition score, and age were significantly (p < 0.05) associated with nematode infestation. Simple logistic regression analysis indicated that nematode infestation in Bena-Tsemay district (78.33%) was significantly (p < 0.05) higher by a factor of 0.54 (OR 95% CI: 0.300.96) than Hamer district (66.39%). Among the species, caprine (79.43%) was significantly (p < 0.05) infested than ovine (62.37%) by a factor of 0.45 (OR 95% CI: 0.250.81). Moreover, infestation on poor (79.12%) and medium (70.96%) body conditioned animals was higher by a factor of 2.94 (OR 95% CI: 1.416.26) and 1.76 (OR 95% CI: 0.883.53) than on good body conditioned animals (63.79%). Likewise, infestation in age groups of 13 years (78.66%) and >3 years (84.40%) was significantly (p < 0.05) higher by a factor of 4.83 (OR 95% CI: 2.3110.46) and 8.23 (OR 95% CI: 3.9817.75) than younger age groups (41.37%), respectively. A moderate parasitic burden was observed on 52.90% of gastrointestinal nematodes (GIN)-infested animals with significantly (p < 0.05) higher EPG in females than males. Furthermore, mixed infestation of Trichostrongylus axei and Eimeria (6.19%), Haemonchus contortus and Eimeria (5.78%), and Trichostrongylus vitrines and Eimeria (5.78%) were dominantly identified. On the contrary, T. axei (15.70%), Eimeria (8.67%), H. contortus (7.43%), and Trichostrongylus colubriformis (7.02%) were dominant single infestations. Conclusion: The current study revealed the highest prevalence of GIN in the study area, which needs strategic control, needs to enhance community awareness toward GIN control and prevention, and to implement further investigation into anthelminthic efficacy to solve the problem. [J Adv Vet Anim Res 2021; 8(4.000): 597-605]

Objective: This case report highlights the first detection of Trypanosoma lewisi, a blood protozoan parasite found in an urban rat in Kota Bharu, Kelantan.  Materials and Methods: Rat trapping was carried out within the Kota Bharu vicinity near a local wet market. A total of 38 rats were captured and subjected to peripheral blood smearing using Giemsa stain. Positive rats were sent for histopathological analysis for the evaluation of the organ samples.  Results: The presence of trypanosomes was found in one sample from a blood smear. This was connected to a histological lesion on kidney tissues, which revealed a high concentration of trypanosomes. Additionally, the positive sample was confirmed as T. lewisi based on molecular diagnosis via polymerase chain reaction and subsequent sequencing and phylogenetic analysis.  Conclusions: This finding serves as a baseline for further surveillance on T. lewisi population among urban rats in Kelantan and possible zoonotic transmission to humans. J. Adv. Vet. Anim. Res., 8(2): 435-439, June 2021 http://doi.org/10.5455/javar.2021.h531

Objective: This case report highlights the first detection of Trypanosoma lewisi, a blood protozoan parasite found in an urban rat in Kota Bharu, Kelantan. Materials and Methods: Rat trapping was carried out within the Kota Bharu vicinity near a local wet market. A total of 38 rats were captured and subjected to peripheral blood smearing using Giemsa stain. Positive rats were sent for histopathological analysis for the evaluation of the organ samples. Results: The presence of trypanosomes was found in one sample from a blood smear. This was connected to a histological lesion on kidney tissues, which revealed a high concentration of trypanosomes. Additionally, the positive sample was confirmed as T. lewisi based on molecular diagnosis via polymerase chain reaction and subsequent sequencing and phylogenetic analysis. Conclusions: This finding serves as a baseline for further surveillance on T. lewisi population among urban rats in Kelantan and possible zoonotic transmission to humans. [J Adv Vet Anim Res 2021; 8(3.000): 435-439]

Objective: The case study describes the cause of an increase in mortality rates among 35-day-old broilers that developed respiratory distress and bloody diarrhea on a farm in Malaysia.  Materials and Methods: The organ samples were subjected to laboratory testing and postmor­tem inspection. Escherichia (E.) coli and Mycoplasma (M.) gallisepticum were detected using bac­terial isolation and molecular diagnostics using polymerase chain reaction.  Results: Chickens with the infection had widespread fibrin buildup in several organs and hemor­rhages on the duodenal mucosa. Additional histology and laboratory analysis of organ samples revealed infection with M. gallisepticum, E. coli, and enteric Eimeria spp., all of which are consis­tent with complex chronic respiratory disease (CCRD) associated with coccidiosis. Tylosin tartrate 20% (w/w) (2.5 gm/l) was prescribed for 1 week along with a combination of the broad-spectrum bacteriostatic drug streptomycin (25 mg/kg) and coccidiostat (2 gm/5 l).  Conclusion: CCRD and coccidiosis are both infectious diseases that can infect chicken flocks, resulting in production losses and carcass quality degradation. Early disease detection and proper treatment should be provided promptly, and tight farm biosecurity should be implemented to prevent chicken mortality on the farm, as was achieved successfully. J. Adv. Vet. Anim. Res., 8(4): 576-580, December 2021 http://doi.org/10.5455/javar.2021.h547

Objective: The case study describes the cause of an increase in mortality rates among 35-day-old broilers that developed respiratory distress and bloody diarrhea on a farm in Malaysia. Materials and Methods: The organ samples were subjected to laboratory testing and postmortem inspection. Escherichia (E.) coli and Mycoplasma (M.) gallisepticum were detected using bacterial isolation and molecular diagnostics using polymerase chain reaction. Results: Chickens with the infection had widespread fibrin buildup in several organs and hemorrhages on the duodenal mucosa. Additional histology and laboratory analysis of organ samples revealed infection with M. gallisepticum, E. coli, and enteric Eimeria spp., all of which are consistent with complex chronic respiratory disease (CCRD) associated with coccidiosis. Tylosin tartrate 20% (w/w) (2.5 gm/l) was prescribed for 1 week along with a combination of the broad-spectrum bacteriostatic drug streptomycin (25 mg/kg) and coccidiostat (2 gm/5 l). Conclusion: CCRD and coccidiosis are both infectious diseases that can infect chicken flocks, resulting in production losses and carcass quality degradation. Early disease detection and proper treatment should be provided promptly, and tight farm biosecurity should be implemented to prevent chicken mortality on the farm, as was achieved successfully. [J Adv Vet Anim Res 2021; 8(4.000): 576-580]

Various types of pain were reported by people with Plasmodium falciparum and were mostly attributed to a symptom of malarial infection. Neural processes of pain sensation during malarial infection and their contributions to malaria-related death are poorly understood. Thus, these form the focus of this study. Swiss mice used for this study were randomly divided into two groups. Animals in the first group (Pb-infected group) were inoculated with Plasmodium berghei to induce malaria whilst the other group (intact group) was not infected. Formalin test was used to assess pain sensitivity in both groups and using various antagonists, the possible mechanism for deviation in pain sensitivity was probed. Also, plasma and brain samples collected from animals in both groups were subjected to biochemical and/or histological studies. The results showed that Pb-infected mice exhibited diminished pain-related behaviours to noxious chemical. The observed parasite-induced analgesia appeared to be synergistically mediated via µ-opioid, α2 and 5HT2A receptors. When varied drugs capable of decreasing pain threshold (pro-nociceptive drugs) were used, the survival rate was not significantly different in the Pb-infected mice. This showed little or no contribution of the pain processing system to malaria-related death. Also, using an anti-CD68 antibody, there was no immunopositive cell in the brain to attribute the observed effects to cerebral malaria. Although in the haematoxylin and eosin-stained tissues, there were mild morphological changes in the motor and anterior cingulate cortices. In conclusion, the pain symptom was remarkably decreased in the animal model for malaria, and thus, the model may not be appropriate for investigating malaria-linked pain as reported in humans. This is the first report showing that at a critical point, the malaria parasite caused pain-relieving effects in Swiss mice.

Various types of pain were reported by people with Plasmodium falciparum and were mostly attributed to a symptom of malarial infection. Neural processes of pain sensation during malarial infection and their contributions to malaria-related death are poorly understood. Thus, these form the focus of this study. Swiss mice used for this study were randomly divided into two groups. Animals in the first group (Pb-infected group) were inoculated with Plasmodium berghei to induce malaria whilst the other group (intact group) was not infected. Formalin test was used to assess pain sensitivity in both groups and using various antagonists, the possible mechanism for deviation in pain sensitivity was probed. Also, plasma and brain samples collected from animals in both groups were subjected to biochemical and/or histological studies. The results showed that Pb-infected mice exhibited diminished pain-related behaviours to noxious chemical. The observed parasite-induced analgesia appeared to be synergistically mediated via µ-opioid, α2 and 5HT2A receptors. When varied drugs capable of decreasing pain threshold (pro-nociceptive drugs) were used, the survival rate was not significantly different in the Pb-infected mice. This showed little or no contribution of the pain processing system to malaria-related death. Also, using an anti-CD68 antibody, there was no immunopositive cell in the brain to attribute the observed effects to cerebral malaria. Although in the haematoxylin and eosin-stained tissues, there were mild morphological changes in the motor and anterior cingulate cortices. In conclusion, the pain symptom was remarkably decreased in the animal model for malaria, and thus, the model may not be appropriate for investigating malaria-linked pain as reported in humans. This is the first report showing that at a critical point, the malaria parasite caused pain-relieving effects in Swiss mice.

In this study, the serological surveillance of Rift Valley Fever virus (RVFV) in southern Egypt was carried out for 460 serum samples collected from domestic animals (unvaccinated), including cattle, sheep, goat, camel and donkey reared in three different provinces (Qena, Luxor and Aswan). Enzyme linked immunosorbent assay (ELISA) was used to detect RVFV antibodies. The results showed that 97 out of 460 animals were positive by using blocking ELISA. The percentage of RVFV infection in cattle, sheep, goat, camel and donkey was 5.55%, 65.21%, 14.44%, 20.65% and 0%, respectively. Geographical distribution and breeding system were taken into consideration for RVFV infection in these animals. The most prevalent type of infection was identified in intensive breeding farms systems (27.63%), and then in individual breeding systems (11.68%). Qena had a higher infection rate of RVFV (23.55%), in comparison to Aswan and Luxor (20.65% and 14.14%, respectively). Marked seroprevalence recorded in this study indicates a high incidence of infection in sheep (65.21%) and camel (20.65%); this necessitates the application of more effective strategies to control these types of infections in Egypt. This study provides a concise picture about the RVFV disease in southern Egypt. We need more similar studies targeted to clarify the reliable epidemiological status of RVFV disease in southern Egypt and other localities.

In this study, the serological surveillance of Rift Valley Fever virus (RVFV) in southern Egypt was carried out for 460 serum samples collected from domestic animals (unvaccinated), including cattle, sheep, goat, camel and donkey reared in three different provinces (Qena, Luxor and Aswan). Enzyme linked immunosorbent assay (ELISA) was used to detect RVFV antibodies. The results showed that 97 out of 460 animals were positive by using blocking ELISA. The percentage of RVFV infection in cattle, sheep, goat, camel and donkey was 5.55%, 65.21%, 14.44%, 20.65% and 0%, respectively. Geographical distribution and breeding system were taken into consideration for RVFV infection in these animals. The most prevalent type of infection was identified in intensive breeding farms systems (27.63%), and then in individual breeding systems (11.68%). Qena had a higher infection rate of RVFV (23.55%), in comparison to Aswan and Luxor (20.65% and 14.14%, respectively). Marked seroprevalence recorded in this study indicates a high incidence of infection in sheep (65.21%) and camel (20.65%); this necessitates the application of more effective strategies to control these types of infections in Egypt. This study provides a concise picture about the RVFV disease in southern Egypt. We need more similar studies targeted to clarify the reliable epidemiological status of RVFV disease in southern Egypt and other localities.