Individual genomic loci, transcript level, and biochemical profile of immune and antioxidant markers associated with genetically identified bacterial mastitis in Shami goats in Egypt.

Background: Mastitis in goats is unquestionably a grave concern, with far-reaching implications for both animal well-being and productivity, while also presenting a potential threat to public health. Aim: The study aimed to compare culture methods and multiplex PCR (m-PCR) in the detection of the most three common mastitis-causing pathogens (Staphylococcus aureus, Escherichia coli, and Streptococcus spp.) and investigate the gene expression, single nucleotide polymorphisms (SNPs), serum concentrations of immunological and antioxidant indicators linked to mastitis in Shami goats. Methods: One hundred Shami do (50 Shami goats with clinical mastitis and 50 normal goats taken as control group). The culture methods and m-PCR analysis were used to find the bacteria in the milk samples. Blood samples were obtained to assess some hemato-biochemical parameters, detect SNPs, and determine the expression of certain immunological and antioxidant indicators in the genes. Results: The culture method detected the pathogens causing mastitis in 90% of the milk samples, but m-PCR detected them in 100% of the milk samples. SNPs linked to mastitis resistance/susceptibility in examined does were detected through DNA sequencing of immunological and antioxidant indicators. The magnitude of gene expression varied significantly between the resistant and mastitis-affected groups. Significant (P ˂ 0.05) elevations were noticed in WBCs count, mainly neutsrophils count, serum levels of BHB, NEFA, triglycerides, LDL-C, AST, ALT, ALP, creatinine, total protein, globulin, Ca, K, GPx, MDA, acute phase proteins, and cytokines in mastitis affected does as compared to control. While RBCs count, PCV%, lymphocytes count, serum concentration of glucose, cholesterol, HDL-C, albumin, Na, Cl, P, GSH, SOD, and catalase significantly (P ˂ 0.05) diminished in mastitis affected does compared to healthy ones. APPs and pro-inflammatory cytokines scored high sensitivities and NPVs but TNF-α and serum amyloid A (SAA) had the highest percentages of increase. Conclusion: The study confirmed that m-PCR is the most sensitive method for bacteria identification (S. aureus, E. coli, and Strept. spp.) while SNPs in antioxidant and immunological genes may be important genetic indicators for mastitis risk or resistance in Shami does. To establish an effective management plan and forecast the most sensitive risk time for illness onset, gene expression profiles of the tested genes may also be employed as proxy biomarkers. TNF-α and SAA may be precious indicators for the detection of caprine mastitis.

Effects of antioxidant vitamins (A, D, E) and trace elements (Cu, Mn, Se, Zn) administration on gene expression, metabolic, antioxidants and immunological profiles during transition period in dromedary camels.

BACKGROUND: Nutrition has a primary role for optimum expression of genetic potential, and most of the farmers have limited resources of green fodder. Hence, a fat-soluble vitamin, especially vitamin A and E and trace elements remained most critical in the animal's ration and affects their productive and reproductive performance adversely. Animals cannot be able to produce these vitamins in their bodies; hence, an exogenous regular supply is needed to fulfil the physiological needs and to maintain high production performance. This study elucidated effects of antioxidant vitamins (A, D, E) and trace elements (Cu, Mn, Se, Zn) administration on gene expression, metabolic, antioxidants and immunological parameters in dromedary camels during transition period. RESULTS: At 0 day, there were no appreciable differences in the expression patterns of the metabolic (IGF-I, ACACA, SCD, FASN, LPL, and BTN1A1) genes between the control and treatment groups, despite lower levels. A substantial variation in the mRNA levels of SOD1, SOD3, PRDX2, PRDX3, PRDX4, PRDX6, and AhpC/TSA was observed between the control and treatment groups, according to the antioxidant markers. In comparison to the control group, the treatment group displayed a significant up-regulation at 0 and 21 days. The treatment and control groups exhibited substantial differences in the mRNA values of IL-1α, IL-1ß, IL-6, and TNFα, as indicated by immunological markers. In comparison to the control group, there was a noticeable down-regulation in the treatment group at 0 and + 21 days. But IL10 produced the opposite pattern. No significant difference was observed in glucose, cholesterol, triglyceride, HDL, total protein, NEFA, BHBA, cortisol and IGF-1 levels between control and treatment group. The activity of serum GPx, SOD and TAC was significantly affected by time and treatment x time in supplemented groups as compared with control group. IL-1, IL-1, IL-6, and TNF were noticeably greater in the control group and lower in the treatment group. Additionally, in all groups, the concentration of all pro-inflammatory cytokines peaked on the day of delivery and its lowest levels showed on day 21 following calving. The IL-10 level was at its peak 21 days prior to calving and was lowest on calving day. CONCLUSION: The results demonstrated a beneficial effect of antioxidant vitamins and trace elements on the metabolic, antioxidant and immunological markers in dromedary camels throughout their transition period.

Single nucleotide polymorphisms, gene expression and serum profile of immune and antioxidant markers associated with postpartum disorders susceptibility in Barki sheep.

The objective of this study was to explore the immunological and antioxidant alterations associated with ovine postpartum disorders. Blood samples were collected from 90 adult Barki ewes and allocated into three equal-sized groups (30 ewes each): control group (CG), inflammatory postpartum disorders group (IPG) and non-inflammatory postpartum disorders group (NIPG). PCR-DNA sequencing approach was carried out for TLR4 (256-bp) and SOD (456-bp) genes, and nucleotide sequence variations were noticed to be associated with postpartum disorders resistance/susceptibility. Gene expression profile was also evaluated and levels of IL5, IL6, IL1-ß, TNF alpha, TLR4 and Tollip were significantly up-regulated in ewes affected with postpartum disorders than resistant ones, while SOD and CAT genes pattern elicited an opposite trend. Exploring serum profile also showed a significant increase of IL-1α, IL-1ß, IL-6, TNF-α, MDA and NO in IPG compared to their correspond values in NIPG and CG. However, serum levels of IL-10, CAT, GSH and GPx were significantly decreased. This study highlights that SNPs in TLR4 and SOD genes could be genetic markers for postpartum disorders resistance/susceptibility in Barki ewes. Gene expression alongside serum profiles of antioxidant markers could also be used to follow-up the immune status of ewes to build up an effective management protocol.

Clinical and Correlated Responses among Steroid Hormones and Oxidant/Antioxidant Biomarkers in Pregnant, Non-Pregnant and Lactating CIDR-Pre-Synchronized Dromedaries (Camelus dromedarius).

Overproduction of free radicals is controlled by antioxidant defense mechanisms. These defense mechanisms are achieved by antioxidant enzymes such as catalase (CAT). The current study aimed to assess the changes in steroid hormones, oxidant/antioxidants biomarkers, lipid profiles/liver functions indices, renal function biomarkers and minerals metabolism in non-pregnant, lactating or pregnant one-humped she-camels (Camelus dromedarius) pre-synchronized with controlled internal drug releasing. The study also focused on the correlational relationships between steroid hormones and the oxidant/antioxidant biomarkers, lipid profiles and liver functions indices, renal functions and mineral metabolism in these she-camels. The study was conducted on apparently healthy dromedary she-camels (n = 60) during breeding season. A sexually active camel-bull was introduced to she-camels pre-synchronized with CIDR. Fifty to sixty days after natural mating, she-camels were examined for pregnancy. She-camels were divided into three main groups according to both pregnancy and lactation as following: pregnant (PREG, n = 38) which was kept as control one, non-pregnant and lactating (LACT, n = 8), and non-pregnant and non-lactating she-camels (NPREG, n= 14). Steroid hormones, i.e., progesterone (P4), estradiol (E2) and cortisol, oxidant indictors, i.e., malondialdehyde (MDA), antioxidant biomarkers, i.e., superoxide dismutase (SOD), total antioxidant capacity (TAC), CAT and reduced glutathione (GSH), lipid profiles indices, renal functions and related minerals were assessed. The present study confirmed the efficacy of using CIDR for synchronization in she-camels. Significant elevations in serum steroids hormones in PREG compare with LACT and NPREG. The highest concentrations of MDA as lipid peroxidation and oxidative stress indictors and lowest levels of antioxidant biomarkers except for SOD, i.e., TAC, CAT and GSH, were reported in PREG compared with LACT and NPREG. PREG showed the highest liver enzymes activities and lowest total protein values. Remarkable increases in serum concentrations of renal function parameters and phosphorous (P) were observed in PREG when compared with the other two groups. The investigated she-camels revealed significant correlation between steroid hormones and the oxidant biomarkers, antioxidant biomarkers, liver functions, renal functions and minerals metabolism parameters. P4 showed positive correlations with antioxidant biomarkers, i.e., TAC, CAT and GSH, and serum aspartate aminotransferase (AST) activities, whereas negative correlations were reported between P4 and renal functions biomarkers, i.e., blood urea nitrogen (BUN), creatinine (Cr) and creatinine kinase (CK), and minerals metabolism parameters, i.e., P and magnesium (Mg), in CIDR pre-synchronized she-camels. In contrast, E2 and cortisol showed negative correlations with antioxidant biomarkers, i.e., TAC, CAT and GSH, lipid profiles/liver functions indices, i.e., AST, alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT), CK and Mg, however, positive correlations were demonstrated between E2 and cortisol, and MDA, Cr and P in investigated she-camels. In conclusion, the present study confirmed the efficacy of using CIDR for synchronization in she-camels. The highest MDA levels as indictors for oxidative stress and the lowest antioxidant levels, i.e., TAC, CAT and GSH, except for SOD in pregnant she-camels, were attributable to physiological oxidative stress as excellent compensatory responses observed in the PREG group to face such a physiologic stage. Moreover, lower P levels in non-pregnant she-camels would be contributed to failure of conception or early embryonic death. The investigated she-camels revealed significant correlations between steroid hormones and the oxidant indicators, antioxidant biomarkers, lipid profile indices and renal functions biomarkers that provided better understanding for physiological stress during pregnancy in camels.

Clinicopathological and bacteriological studies on lamb bacterial enteritis and monitoring the oregano oil and vitamins A,D3,E effect on its treatment.

OBJECTIVE: The objective of the study was to assess the effect of A,D3,E (I/M) and oregano oil extract 15% on some clinicopathological parameters during lamb bacterial enteritis treatment. MATERIALS AND METHODS: Sixty Barki lambs, 20 apparently healthy (control group) and, 40 suffered from bacterial enteritis [enteric group (EG)], were subdivided into four treated groups (TGs): antibiotic group (AG), antibiotic + A,D3,E group (A + A,D3,E), antibiotic + oregano oil (AOG), and oregano group (OG). Fecal swabs were collected from EG then aseptically cultured, isolated, phenotypically identified, genotypically confirmed, and sequenced by PCR 16srRNA. Paper disk diffusion test was used for estimation of oregano oil extract 15% antibacterial activity. After blood sample aspiration from all animals, they were clinicopathologically and statistically analyzed. RESULTS: Escherichia coli, followed by Salmonella species and then Klebsiella species, was the main causative agents of lamb diarrhea and were susceptible to oregano oil extract 15%. A + A,D3,E and AOG showed significant (p < 0.05) enhancement of some clinicopathological parameters more than AG or OG. Matrix metalloproteinases (MMP-2 and MMP-9) and total antioxidant capacity (TAC), yielded area under the curve, sensitivity, negative predictive value as 1, 100% and 100% respectively, were determined in both EG and TGs. CONCLUSION: Oregano oil extract 15% has good antibacterial properties against enteric bacteria in vitro and in vivo. The combination between antibiotic and antioxidant vitamins or oregano plant extract of 15% has a good impact on some clinicopathological alterations in lamb bacterial enteritis treatment. TAC, MMP-9, and MMP-2 may be good markers for the disease and its treatment follow-up.

The effects of saline water consumption on the ultrasonographic and histopathological appearance of the kidney and liver in Barki sheep.

The objective of this study was to evaluate the impact of varying degrees of water salinity on the ultrasonographical and histopathological appearance of the liver and kidneys in Barki sheep. Thirty Barki sheep (initial weight, 29.48 ± 0.81 kg) were allocated into three groups (n=10 per group) based on the type of drinking water for 9 months: the tap water (TW) group (350 ppm total dissolved solids [TDS]); the moderate saline water (MSW) group (4,557 ppm TDS); and the high saline water (HSW) group (8,934 ppm TDS). After 9 months, the body weight was significantly decreased in sheep subjected to MSW (P=0.0347) and HSW (P=0.0424). Alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, urea, and creatinine were significantly increased (P<0.05) in sheep subjected to MSW and HSW. Ultrasonographic examination of the right and left kidneys revealed an increased length of both kidneys with crystal formation, particularly in male sheep. Ultrasonographic examination of the liver showed hyperechogenic dots varying in size and number between males and females. Histopathological examination of kidney revealed significant changes in both MSW and HSW groups such as hyaline matrix formation, atrophied glomerular tufts, and intramedullary congestion. Histopathological examination of the liver revealed slight fatty liver changes, slight fibrosis around the bile duct, massive inflammatory cell infiltration and vacuolar changes of hepatocytes in both MSW and HSW groups. In conclusion, water salinity negatively affects the body weight, liver and kidney appearance of Barki sheep and thus sheep production.