Ticks transmit a number of bacterial, protozoal and viral pathogens that cause many diseases like ehrlichiosis, hemorrhagic fever, theileriosis, babesiosis and anaplasmosis in livestock. This study was designed for molecular characterization of BM86 gene of Rhipicephalus (Boophilus) microplus tick. In this study, the BM86 gene was amplified, using primers fl anked by restriction enzyme sites. The molecular detection of R. microplus was studied in three districts belonging to diff erent ecological zones in the province of the Punjab, Pakistan. Tick samples were collected and initially screened through microscopy and further analyzed by PCR and sequencing. The phylogenetic tree was generated by using the MEGA 7 through Neighbor Joining method employing best model through the phylogenetic analysis of R. microplus. Pairwise comparisons of nucleotide sequences showed nucleotide diff erences ranging between 0.007 and 0.01%. Haplotype and nucleotide diversity in Bm86 gene was found among diff erent districts. Six single nucleotide polymorphisms were seen in sequences of BM 86 from indigenous tick populations collected from the Punjab province. More interestingly, out of these 6 polymorphisms, we got 2 from district Okara, 3 from district Sahiwal, and 1 in Mandi Bahauddin. Conserved regions were observed among the local strains for BM86 gene. A common convergence in similar clade was with the local Pakistan R. microplus. Local mean diversity was 0.005 and overall mean diversity was 0.038. Field strain has been isolated as candidate specie for local tick vaccine, which in turn will increase the efficacy of future tick vaccine including reduction of economic burden on the farmer.
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The aim of the present study was to investigate the potential protective eff ects of Chitosan (C) and Chitosan Oligosaccharide (COS) on sodium fl uoride (NaF) induced testicular damage. Forty-two male Wistar rats were separated into six groups (n=7). The Control group was given drinking water without Fluoride. NaF group was given NaF (100 mg/L) in their drinking water. NaF+C group was given NaF (100 mg/L) and C (250 mg/kg/day). NaF+COS group was given NaF (100 mg/L) and COS (250 mg/kg/day). C group was given C (250 mg/kg/day). COS group was given COS (250 mg/kg/day). Rats were given C and COS by gastric gavage. As a result, the total number of spermatogonia, Leydig cells, the total volume of the testis, the total volume of germinal epithelium, the total volume of seminiferous tubules, the total length of seminiferous tubules, and the height of germinal epithelium significantly decreased in the NaF group compared to the control group (P<0.05). These parameters significantly increased in the NaF + C and NaF + COS groups compared to the NaF group (P<0.05). Also, GSH and CAT significantly decreased, while MDA significantly increased in the NaF group. C and COS alleviate these changes. These findings indicate that NaF can cause testis damage. C and COS can have cytoprotective eff ects against testis damage
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In humans and animals, botulism is a disease characterized by generalized and progressive paralysis caused by Clostridium botulinum neurotoxins (BoNT). BoNTs, defined in seven diff erent antigenic types (A to G), proteolyze SNAREs (synaptosomal-associated protein/SNAP-25 and synaptobrevin) responsible for acetylcholine release in peripheral cholinergic neurons, and thus cause fl accid paralysis and death. Currently, mouse experiments are considered the reference method for definitive diagnosis. However, new diagnostic methods that are fast and accurate and would not raise ethical issues need to be developed. Therefore, using antibodies specific to the toxoid forms of BoNTs, the presence of BoNT-C and/or BoNT-D was investigated by immunohistochemical method (IHC) in the study. The tissues of thirty ruminants (twenty cattle, seven sheep, three goats), which had the clinical and pathological findings of botulism and a herd history of the disease, were used as material. BoNTs were detected with IHC in sixteen of the thirty ruminants as three BoNT-C, eleven BoNT-D, and two BoNT C+D. In the mouse experiments, BoNT was isolated in only three cases (two BoNT-D, one BoNT-C). Additionally, being responsible for the clinical findings of botulism, the interaction of BoNTs with SNAP-25 and synaptobrevin was investigated using IHC. It was determined that BoNT-C specifically reduces the expression of SNAP-25, and BoNT-D reduces the expression of synaptobrevin and partially SNAP-25. It was concluded that additional studies may be valuable to investigate the use of IHC in the diagnosis of botulism.
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The unique geographical and ecological conditions of the Qinghai-Tibetan Plateau (QTP) constitute the environmental conditions of the natural foci of cystic echinococcosis (CE). It is a rare and highly endemic area in the world. The main host of the pathogen, Tibetan sheep, is widely distributed, and the population density is on the rise in recent years, which increases the pressure on the transmission and risk of the natural epidemic source. Currently, the developments of serological diagnosis test for Echinococcus granulosus infection on the QTP Tibetan sheep are scanty, which seriously restricts the epidemiological investigation of CE in Tibetan sheep on the QTP. EgAgB8/2 is one of the most important targets of host antibody response in CE and has better diagnostic performance in livestock. This study expressed the recombinant EgAgB8/2 antigen of E. granulosus from Tibetan sheep in prokaryotic expression vector, and also preliminarily evaluated its potential value for diagnosing CE in Tibetan sheep using indirect ELISA. Our preliminary results shown that the recombinant EgAgB8/2 antigen had good immunogenicity and exhibited high sensitivity (95%) and high specificity (100%) and no cross-reacted with both Taenia multiceps and Taenia hydatigena. Further studies are needed to collect more Tibetan sheep sera from E. granulosus and other parasitic infections, which may evaluate further sensitivity, specificity and cross-reactive of recombinant EgAgB8/2 antigen in Tibetan sheep positive for CE of the QTP, China.
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Transglutaminase (TGase) is an enzyme widely used in the food industry. In this study, the eff ect of transglutaminase enzyme on the chemical and physical characteristics of cooked beef meatballs was evaluated. For this aim, beef meatballs were prepared by using recombinant microbial transglutaminase (MTGase) and commercial TGase enzymes, after that physical and chemical tests were applied to meatball samples. The addition of MTGase enzyme improves the quality parameters of the beef meatballs. The myofibrillar proteins of cooked meatball samples were also analyzed with SDS-PAGE analysis. It was observed that, actin and myosin proteins bind covalently to form a new high molecular weight protein by the help of MTGase addition. These results indicated that recombinant MTGase enzyme can be used to obtain high quality restructured beef meat products.
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Zinc transporters (ZnT) and ZIP proteins maintain Zinc homeostasis in the live organisms. On the other hand, the impacts of zinc oxide nanoparticles (ZnO NPs) on the expression of the Znt genes in biological systems were not clear yet. So that in this experimental study we have tried to find the eff ects of ZnO NPs on Znt1-4 genes expression in the hippocampus of male rats under acute stress. Adult male rats were divided into groups of control and treated with 5 or 10 mg/kg of ZnO NPs alone and under acute restraint stress for 90 min. The changes in the expression of the selected genes were monitored using real-time qRT-PCR. The ZnT4 protein expression also was measured by Western blotting. Real-time qRT-PCR expression analysis revealed that the Znt1 gene expression was up-regulated in the stress group, while the expression of the Znt1 and Znt4 genes was significantly up-regulated in the group receiving 10 mg/kg of ZnO NPs. Furthermore, in the ZnO NPs 10 mg/kg group under stress, the Znt2 gene expression was down-regulated, while the Znt4 gene expression was up-regulated. Moreover, the levels of ZnT4 protein were significantly increased after 10 mg/kg of ZnO NPs injection in the stress and normal groups. According to these results ZnO NPs administration can cause changes in the expression of a number of zinc transporter genes under stress conditions and increases the ZnT4 protein level. Therefore, this is a valuable approach for forecast investigation in biomedicine and pharmacogenetics studies.
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Newcastle Disease (ND) was ranked as a List A and trade limited disease by the World Health Organization. It is one of the deadly diseases of domestic and zoo birds especially pheasants that are highly susceptible species to Newcastle disease virus (NDV). This study was designed to determine the humoral immune response (HIR) of thermostable NDV vaccine strain I-2 in same age pheasants. For this purpose, forty-five pheasants of the same age were separated and placed in cages. Thermostable NDV I-2 vaccine was mixed with feed and administered through oral route to the same age pheasants. HIR was detected using haemagglutination inhibition test (HI) and enzyme linked immunosorbent assay (ELISA) on 0, $7^{th}$, $14^{th}$, $21^{th}$ and $28^{th}$ days post-vaccination (DPV). Optimum geometric mean anti-NDV-ELISA (2380) and anti-NDV-HI (Log27.5) antibodies titers were identified on $14^{th}$ DPV. It was concluded that the oral administration of NDV I-2 strain is able to elicit a protective immune response in pheasants. Moreover, the use of this novel vaccine technique at the same age of pheasants overwhelms the attempt to catch these birds for single vaccination.
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Lower urinary tract dysfunction (LUTD) is a commonly seen problem in cats. This chronic condition with no specific underlying cause remainsa challenge for achieving eff ective treatment. Glycosaminoglycans (GAGs) are linear polysaccharides possessing characteristic repeateddisaccharide sequences, thought to be involved in the pathogenesis of feline LUTD. The aim of the present study is to evaluate feline-specificserum sulphated glycosaminoglycan (S-GAG) and serum dermatan sulphate (DS) levels in cats with non-obstructive LUTD versus healthycontrols. Eighteen client-owned cats suff ering non-obstructive LUTD and 16 client-owned healthy cats were enrolled in this case-controlstudy. Pre-treatment serum samples from cats in both the study (non-obstructive LUTD cats) and control (healthy cats) groups were analysedwith “Quantitative Sandwich ELISA method” using feline-specific S-GAG and DS kits. The mean serum S-GAG and DS levels of the study groupwere measured against the control group. Measurements for study males were compared to the control males, and the neutered cats inthe study group were compared to the intact ones in control group. Cats in study group had lower serum S-GAG concentrations (3.52±0.26ng/mL) than the control ones (3.93±0.27 ng/mL). Cats in study group had higher serum DS levels (27.20±6.62 ng/mL) than control cats(16.79±5.21 ng/mL). This study reports serum S-GAG and DS data in cats with non-obstructed LUTD and in healthy cats for the first time.
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Glutamate is the major excitatory neurotransmitter in the central nervous system. Excessive glutamate is known to cause excitotoxicity.Umbelliferone is a coumarin derivative compound and has antioxidant, anti-infl ammatory, and neuroprotective eff ects. Also, umbelliferone can showneuroprotective eff ect by crossing the blood-brain barrier. In our study, it was aimed to investigate the neuroprotective eff ect of umbelliferone onprimary cortical neuron (PCN) culture. Umbelliferone was isolated from the roots of Ferulago cassia dichloromethane sub-extract. The cerebral cortexof newborn Sprague Dawley rats was used to obtain PCNs. To stimulate glutamate excitotoxicity, cells were exposed to 6x10-5M glutamate. Thendiff erent concentrations (10-1000 μM) of umbelliferone were added into the medium and allowed to incubate for 24 and 72 h. MTT assay was used tomeasure cell viability. Total Antioxidant Status (TAS) and Total Oxidant Status (TOS) analyzes were used to evaluate reactive oxygen species. MTT resultsshowed that cell viability was decreased with glutamate application. 25-250 µM umbelliferone had a significant protective eff ect against glutamateexcitotoxicity at 72 h (P<0.05). Consistent with MTT results, TAS analysis results showed 50-250 µM umbelliferone increase the level of antioxidantsin cells, which can help protect neurons against glutamate-induced excitotoxicity. In this study, umbelliferone showed a neuroprotective eff ect inPCN against glutamate excitotoxicity. These results suggest that umbelliferone may be used as therapeutic agent against glutamate excitotoxicity.
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The purpose of this study was to investigate the effi cacy of synovial fl uid (SF) as a source of hyaluronic acid (HA) on Limberg and Elliptical rotationfl ap healing. In this study conducted on 24 male Mus musculus mice, the animals were divided into 2 main groups. Group 1 (n = 12): Limbergfl apgroup; Group 2 (n = 12) Elliptical rotation fl ap group. Six of the animals in Group 1 and Group 2 were assigned as the Flap (Control) Group, and 6 asthe Flap + SF Group. In SF subgroups of animals in each group, 1 mL SF was injected into the operation area in addition to the fl ap surgical procedure.The groups were compared in terms of epithelialization, angiogenesis, neutrophil leukocyte infiltration and fibroblast activity, and the eff ect of SFon the infl ammation and wound healing was evaluated. It was observed that epithelization was completed in all animals in the groups which wereadministered compared to the other groups, and there was a statistically significant diff erence (P<0.05). In addition, no necrosis or abscess wasfound in the groups which were administered SF. There was no statistical diff erence in the paired comparisons of neutrophil-leukocyte infiltrationand fibroblast activity parameters. Although the Limberg fl ap technique is more commonly used today, it can be said that elliptical rotation fl ap isa method that can be used in routine practice due to a lower complication rate. However, SF increases epithelization and plays a regulatory role onangiogenesis. Therefore, it is concluded that the use of SF as a source of HA in combination with elliptical rotation fl ap in wounds with material tissueloss will contribute to clinical practice.
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