Isolated Bacteria from the Uteri of Camels with Different Reproductive Backgrounds: A Study on Sampling Methodology, Prevalence, and Clinical Significance.

The objectives of this study were to comparatively identify the common bacterial isolates from the uteri of camels coming from different reproductive backgrounds after standardizing the sampling method and to investigate the association of clinically measurable parameters with uterine colonization by these isolates. The uterine samples from 856 dromedary camels yielded a total of 17 different bacterial species with a higher proportion of sub-fertile camel uteri being colonized by bacteria (66.6%) as compared to nulliparous, recently calved, and those with unknown reproductive history combined (44.2%; p < 0.05). Camels with body condition scoring < 3 and those with a consistently echogenic appearance of the uterine lumen by sonography were more likely to be positive on uterine culture, while the presence of pus in uterine discharge was not associated with the odds of bacterial isolation (p > 0.05). While certain strains were more likely to be obtained from the uteri of the sub-fertile group (p < 0.05), embryo transfer to camels with a positive uterine culture in the absence of other gross reproductive pathologies did not necessarily affect the overall pregnancy rate compared to recipients with a negative uterine culture (p > 0.05). In conclusion, a relatively high bacterial load can be identified from the uteri of both sub-fertile and normal dromedary camels, with a higher frequency among the former. The uterine ultrasonography and evaluation of the body condition score can help in identifying the camels in which uterus is contaminated by bacteria.

Isolation of infectious laryngotracheitis virus in broiler chicken in Iran: First report.

In February 2019, a severe respiratory distress with co-infection of infectious laryngotracheitis (ILT) and Newcastle disease accompanied with Salmonella Enteritidis occurred in a broiler flock in the western region of Iran. Clinical signs included paralysis, torticollis, nasal discharge, conjunctivitis, gasping and respiratory rale with high mortality. At necropsy, caseous diphtheritic membrane adherent to the larynx and trachea was observed. Microscopically, syncytial cells formation with dense eosinophilic intranuclear inclusion bodies were main histopathological findings in tracheal tissues. Conventional polymerase chain reaction (PCR) for ICP4 gene amplification as a definitive diagnosis was utilized for the detection of ILT virus nucleic acid in suspected tracheal samples inoculated on to the chorioallantioc membrane of 11-day-old specific pathogen free (SPF) chicken eggs. Tracheal tissues taken from these SPF birds were positive by nested ILT PCR. In conclusion, because of no vaccination policy against ILT in broilers, the most probable scenario is that virus-laden dust or other fomites can be vectors and virus persistence and disease outbreak can be a sequel of wild virus introduction to the farm.

Coronavirus: proteomics analysis of chicken kidney tissue infected with variant 2 (IS-1494)-like avian infectious bronchitis virus.

Avian infectious bronchitis virus is one of the most important gammacoronaviruses, which causes a highly contagious disease. In this study, we investigated changes in the proteome of kidney tissue of specific-pathogen-free (SPF) chickens that were infected with an isolate of the nephrotropic variant 2 genotype (IS/1494/06) of avian coronavirus. Twenty 1-day-old SPF White Leghorn chickens were randomly divided into two groups, each comprising 10 chickens, which were kept in separate positive-pressure isolators. Chickens in group A served as a virus-free control group up to the end of the experiment, whereas chickens in group B were inoculated with 0.1 ml of 104.5 EID50 of the IBV/chicken/Iran/UTIVO-C/2014 isolate of IBV, and kidney tissue samples were collected at 2 and 7 days post-inoculation (dpi) from both groups. Sequencing of five protein spots at 2 dpi and 22 spots at 7 dpi that showed differential expression by two-dimensional electrophoresis (2DE) along with fold change greater than 2 was done by MS-MALDI/TOF/TOF. Furthermore, the corresponding protein-protein interaction (PPI) networks at 2 and 7 dpi were identified to develop a detailed understanding of the mechanism of molecular pathogenesis. Topological graph analysis of this undirected PPI network revealed the effect of 10 genes in the 2 dpi PPI network and nine genes in the 7 dpi PPI network during virus pathogenesis. Proteins that were found by 2DE analysis and MS/TOF-TOF mass spectrometry to be down- or upregulated were subjected to PPI network analysis to identify interactions with other cellular components. The results show that cellular metabolism was altered due to viral infection. Additionally, multifunctional heat shock proteins with a significant role in host cell survival may be employed circuitously by the virus to reach its target. The data from this study suggest that the process of pathogenesis that occurs during avian coronavirus infection involves the regulation of vital cellular processes and the gradual disruption of critical cellular functions.

The effect of Allium sativum (Garlic) extract on infectious bronchitis virus in specific pathogen free embryonic egg.

OBJECTIVE: Garlic is a plant has been used as a flavor, and anti-microbial and anti-diarrheal agent. Infectious bronchitis virus (IBV) is a coronavirus. The available vaccines against IBV cannot cover new variants. This study evaluated the inhibitory effects of garlic extract on IBV. MATERIALS AND METHODS: The constituents of garlic extract were detected by gas chromatography. This study was done in four groups of embryonic SPF eggs; first group was used for virus titration; second group received the mixture of different virus titration and constant amount of garlic extract; third group received 10(-3) titration of virus and after 8 hr received garlic extract and the last group received different dilutions of garlic extract. RESULTS: Based on our results, in the second group, IBV vaccine strain (4/91) at all titration and M41 in 10(-2) and 10(-3) titration and in the third group both variants of virus the embryonic Index (EI) was significantly increased. CONCLUSION: The garlic extract had inhibitory effects on IBV in the chickens embryo.

Paracrine Neuroprotective Effects of Neural Stem Cells on Glutamate-Induced Cortical Neuronal Cell Excitotoxicity.

PURPOSE: Glutamate is a major excitatory neurotransmitter in mammalian central nervous system. Excessive glutamate releasing overactivates its receptors and changes calcium homeostasis that in turn leads to a cascade of intracellular events causing neuronal degeneration. In current study, we used neural stem cells conditioned medium (NSCs-CM) to investigate its neuroprotective effects on glutamate-treated primary cortical neurons. METHODS: Embryonic rat primary cortical cultures were exposed to different concentrations of glutamate for 1 hour and then they incubated with NSCs-CM. Subsequently, the amount of cell survival in different glutamate excitotoxic groups were measured after 24 h of incubation by trypan blue exclusion assay and MTT assay. Hoechst and propidium iodide were used for determining apoptotic and necrotic cell death pathways proportion and then the effect of NSCs-CM was investigated on this proportion. RESULTS: NSCs conditioned medium increased viability rate of the primary cortical neurons after glutamate-induced excitotoxicity. Also we found that NSCs-CM provides its neuroprotective effects mainly by decreasing apoptotic cell death rate rather than necrotic cell death rate. CONCLUSION: The current study shows that adult neural stem cells could exert paracrine neuroprotective effects on cortical neurons following a glutamate neurotoxic insult.

Full-length characterization and phylogenetic analysis of hemagglutinin gene of H9N2 virus isolated from broilers in Iran during 1998-2007.

H9N2 avian influenza A viruses (AIV) have become panzootic in Eurasia over the last decade and are endemic in Iran since 1998, and inactivated vaccine has been used in chickens to control the disease. The hemagglutinin (HA), one of eight protein-coding genes, plays an important role during the early stage of infection. To study their evolution and zoonotic potential, we conducted an in silico analysis of H9N2 viruses that have infected broiler in Tehran Province, Iran between 1998 and 2007. The complete coding region of HA genes from nine H9N2 subtypes isolated from chicken flocks in Tehran Province during 1998-2007 was amplified and sequenced. Sequence analysis and phylogenetic studies of H9N2 subtype viruses on the basis of data of 9 viruses in this study and 30 selected strains are available in the GenBank. Sequence and phylogenetic analyses revealed a large number of similar substitution mutations and close evolutionary relation among sequences of HA. The isolates possessed two types of amino acid motif -R-S-S-R/G-L- and -R-S-N-R/G-L- at the cleavage site of HA. The results showed that all nine representative H9N2 isolates belong to low pathogenic AIVs since none of the amino acid sequences at the cleavage site of the HA of the isolates possessed the basic motif required for highly pathogenic viruses (R-X-R/K-R). Six out of these nine isolates possessed leucine at position 226, which prevails in the sequences found in human strains. Phylogenetic analysis showed that all our isolates belonged to the G1-like sublineage. Also, these isolates showed some degree of homology with other H9N2 isolates, e.g., 89.46-93.93.39% with qu/HK/G1/97 and 93.39-98.39% with pa/Narita/92A/98. The available evidence indicates that HA genes of H9 influenza virus circulating in Iran during the past years were not well conserved. Our finding emphasizes the importance of reinforcing AIV surveillance, especially after the emergence of high pathogenicity in poultry in Iran.

Cell damage through pentose phosphate pathway in fetus fibroblast cells exposed to methyl mercury.

Methylmercury (MeHg) is a global pollutant that causes malformations. There has been no direct evidence for the effect of MeHg on pentose phosphate pathway (PPP). In embryonic development, PPP is much more active. This pathway produces ribose for DNA/RNA production. It is possible that one of teratogenicity mechanisms of MeHg is through PPP. The fetus fibroblast cells were incubated with different concentrations of MeHg (0.1-100 µm). A dose-response dependence was observed in MTT assay. Transketolase activity and DNA content were determined in cell exposed to MeHg. A defect at the level of DNA content was observed. This amount of DNA was highly correlated with transketolase activity (r = 0.76). This study has demonstrated that the potential teratogenic action of MeHg is through PPP. To assess the protective effects of thiamin, the infected cells were incubated with different concentrations of thiamin. The obtained results show that thiamin pyrophosphate supplementation correlated with the toxicity. This finding confirms that thiamin therapy is suitable for the prevention of MeHg toxicity. Our study provides basic data for prevention and treatment of MeHg toxicity via boosting PPP.

Efficacy of conventional and extended intra-mammary treatment of persistent sub-clinical mastitis with cefquinome in lactating dairy cows.

The objectives of the present study were to evaluate the efficacy of intra-mammary-administered cefquinome for the treatment of sub-clinical mastitis in lactating dairy cows and to determine if extended therapy would enhance treatment efficacy. Seventy-three Holstein dairy cows from a single farm with 150 infected quarters were enrolled in the study. Infected cows were allocated randomly to one of three treatment regimens: (1) conventional (standard) regimen: 75 mg of cefquinome administered three times at 16-h intervals (25 infected cows, 52 intra-mammary infections (IMI)), (2) extended regimen: 75 mg of cefquinome administered six times at 16-h intervals (26 infected cows, 58 IMI) and (3) negative untreated control group (22 cows, 40 IMI). Most IMI were caused by coagulase-negative staphylococci, streptococci other than Streptococcus agalactiae and coliforms. The overall bacteriological cure (BC) rates for sub-clinical IMI were 84.61%, 91.37% and 20% for the conventional, extended and the control groups, respectively, indicating a higher BC rate for the treated groups than the control group (P < 0.001). Significant differences in somatic cell count (SCC) were detected between the treated versus the control group (P < 0.001). No differences, concerning the BC rate or SCC, were observed between the extended and the conventional groups. Although fat and protein percentages increased in the treated groups, there were no significant differences in post-treatment milk production between the groups. Results of this study indicate that cefquinome therapy was effective in reducing SCC and eliminating sub-clinical IMI in lactating dairy cows, but extended therapy did not enhance treatment efficacy.

Biological screening of Amaranthus retroflexus L. (Amaranthaceae) / Abordagem biológica de Amaranthus retroflexus L. (Amaranthaceae)

Bioassays are required for the determination of the total toxicity of Amaranthus retroflexus L. (Amaranthaceae) or "redroot pigweed". Therefore, the plant extract has been tested for bioactivity in Artemia salina and cytotoxicity against bovine kidney cells. The LD50 values for Artemia salina were measured at 1700 ppm. The bovine kidney cells were exposed to various concentrations of the plant extracts (100 ppm-0.1 ppm). After treating with 100 and 0.1 ppm for 24 h, the cells viability were reduced by about 49 percent and 35 percent respectively in MTT viability assay. The study confirmed that Amaranthus retroflexus has a cytotoxic effect and more specific to renal cells.

Ensaios biológicos foram realizados para determinação da toxidade de Amaranthus retroflexus L. (Amaranthaceae) conhecido popularmente como "redroot pigweed". Extratos desta espécie foram testados para avaliar sua bioatividade em Artemia salina e citoxidade em células bovinas de rim. Os valores de DL50 para Artemia salina foram medidos a 1700 ppm. As células de rim bovinas foram expostas a várias concentrações dos extratos de plantas (100-0,1 ppm). Após tratamento com 100 e 0,1 ppm por 24 h, a viabilidade celular foi reduzida a cerca de 49 por cento e 35 por cento, respectivamente, no ensaio de MTT. O estudo confirma que Amaranthus retroflexus apresenta um efeito citotóxico e, mais especificamente, para células renais.

Histopathology and cholinergic assessment of Pterocarya fraxinifolia on chicken embryo.

There are no reports of toxicological studies of Pterocarya fraxinifolia. The leaves are used for fishing, which also an anesthetic agent. Currently, many drugs utilized in anesthesia practice are modified cholinergic transmission and acetylcholine esterase inhibitors; these are parts of anaesthetic pharmacy. Therefore, cholinergic assessment was surveyed in chicken embryo, which Pterocarya fraxinifolia extractes were injected in 0.1, 1 and 10 mg concentration at day 4 of incubation. Serum and brain cholinesterase were analyzed on day 20 of incubation. The signs were not due to the changes of cholinesterase activity. In histopathology examination, massive necrosis was observed in the spinal cord. Other tissues such as heart, kidneys, skeletal bones and muscles, trachea and lungs, digestive system and endocrine glands were completely developed. This data suggests that the spinal cord is a target organ of the bioactive component of this plant.