Iranian Journal of Veterinary Research
Volume:20 Issue: 2, Spring 2019

Zataria multiflora Boiss. (ZM) from Lamiaceae family, is an herb native to Iran, Pakistan, and Afghanistan with a history of use both as a condiment and a traditional medicine. The herb and/or its preparations have also shown promising effects in modern pharmacological studies. With regard to the importance of reducing synthetic drug administration both as growth promoters and therapeutic agents in poultry industry, this review is a short account of current knowledge about the beneficial effects of this herb on broiler health and performance parameters based on published materials in different databases such as Google Scholar, PubMed, ScienceDirect, Scientific Information Database, Scopus, etc. It seems that ZM, possibly due to the presence of thymol and carvacrol as its main constituents, can affect broiler’s health especially with regard to its antioxidant effects, immunomodulatory properties and proper effects on gut bacterial pathogens and/or microbiota. It should be kept in mind that although major constituents of ZM essential oil (as the most studied preparation of the herb) are relatively similar, their relative content may show a broad range that can subsequently affect the outcome of ZM essential oil administration to broilers. Overall, ZM is a pluripotent herb and its effect on broilers should be more clarified in future studies.

The sub-clinical carry-over effects of post-partum (PP) conditions and the problems independent of parturition may affect the cows’ performance during the breeding period. It was hypothesized that some mid-lactation cows may have compromised liver functions and calculating liver activity index (LAI; -1.5 to +1.5) may be helpful in detecting such conditions. Plasma lipid and protein profiles, retinol and ceruloplasmin were measured in 37 Holstein cows on days 60, 90, and 120 PP. Liver activity index was calculated using the measures of cholesterol, albumin and retinol. The results were interpreted using some health and performance indices. The mean LAI was 0.00 ± 0.61. Sixteen cows (45.7%) were LAI‾ and 19 cows (54.3%) were LAI+. In LAI+ cows the concentrations of cholesterol and albumin were unchanged, but in LAI‾ cows both of them increased through the study (P<0.05). Greater concentrations of low density lipoproteins (LDL) (P<0.01) and lower concentrations of γ- and total globulins (P<0.05; day 60) were detected in LAI+ cows. Ceruloplasmin was not different between the LAI groups with the highest level on day 90. Cholesterol had the strongest and the most repeated correlations with LAI during the study. The correlation of albumin with LAI faded on day 120 and vitamin A had a relationship only on day 60 PP. Triglycerides (TGs), very low density lipoproteins (VLDL) (day 60) and LDL (days 60 and 90) had positive correlations with LAI. All globulin fractions showed negative correlations with LAI on day 60. Seventy percent of the cows without endometritis (day 30 PP) and 33% of the infected cows were LAI+ during the breeding period. About 79% of the cows with body condition score (BCS) loss ≤0.75 (day 60) and 38% of the cows with BCS loss >0.75 were LAI+ during the breeding period. Compromised liver functions may exist in dairy cows during the breeding period and may be detected by calculating LAI. The relationship of LAI during the progressed lactation with herd’s performance needs to be investigated.

One of the factors limiting successful processing of alpaca (Vicugna pacos) semen is the viscosity of seminal plasma. The viscous nature of the collected ejaculate has hindered sperm cryopreservation as well as artificial insemination (AI) under field conditions. The objective of this investigation was to evaluate recovery, motility, and plasma membrane integrity of alpaca spermatozoa after centrifugation in one of two different solutions at one of three different combinations of speed and time. A total of 24 ejaculates was recovered from seven reproductively sound Huacaya males using a modified artificial vagina (AV) after training the animals for semen collection. A 2 × 3 factorial treatment arrangement was utilized for this study. Ejaculates were divided into fractions for centrifugation in one of two solutions (Tris extender or PureSperm®80 density gradient solution) at one of three combinations of speed and time (492 × g for 15 min, 1968 × g for 10 min, or 4448 × g for 7 min). The experiment was replicated eight times. Analysis revealed that centrifugation at 4448 × g for 7 min in PureSperm80 provided a high recovery rate of spermatozoa with the highest sperm motility and functional integrity of plasma membrane post-centrifugation. Results suggest that adoption of this procedure (centrifugation at 4448 × g for 7 min in PureSperm80) in the initial processing of alpaca ejaculates may enhance subsequent ability to use semen for AI and other assisted reproductive biotechnologies in this species.

Avian reovirus (ARV) has a global distribution in nature and most clinical signs are found in broiler type chickens. This study was conducted to detect and identify reovirus infections from vaccinated breeder chickens and their progenies. A total of 20 tissue and blood samples were collected from vaccinated broiler breeders and their progenies with gastrointestinal or performance problems during peak production. Antibody titers were measured by indirect enzyme-linked immunosorbent assay (ELISA) tests. RNA extraction from tissue samples was performed and cDNA was prepared and directly used in the polymerase chain reaction (PCR). Nucleotide sequences were bilaterally determined using internal primers. The analysis of the nucleotide sequences and their related amino acids was performed by the specialized Molecular Evolutionary Genetics Analysis software (6th version). The virus variant was detected in two vaccinated broiler breeders and five broiler flocks. The vaccine strains in breeder flocks included S1133, SS412, 1733, 2408 belonging to genotype 1 from the reovirus phylogenetic tree. Sequence 7 from the isolated reovirus based on the σC revealed that they were different from the reovirus vaccine, and that 6 isolates belonged to genotype 1 of the phylogenetic tree while 1 isolate belonged to branch 4 of the phylogenetic tree. The study showed that the new reovirus strain isolated from vaccinated birds differs from common strains used in the vaccines. It is therefore essential to prevent the effects of the field reovirus on the performance of industrial poultry, by updating and inventing new commercial vaccines, live and killed, against the reovirus.

Anthrax is a particularly dangerous infectious disease that affects humans and livestock. Efficacious vaccines that can rapidly induce a long-term immune response are required to prevent anthrax infection in humans. Domains 4 and 1 of the protective antigen (PA) and lethal factor (LF), respectively, have very high antigenic properties. In this experimental study, the pET28a-lfD1-pa4 expression vector was designed, constructed and transferred into E. coli BL21 (DE3) plysS. For this purpose, pa4 gene was amplified by polymerase chain reaction (PCR) and cloned in a pGEM T-easy vector. The pGEM-pa4 and pGEM-lfD1 were digested by XbaI and HindIII enzymes. The ligation reaction was performed by ligase T4 enzyme and the gene cassette, lfD1-pa4, was subcloned in pET28a and transferred to E. coli BL21 (DE3) PlysS. Expression and purification of chimeric proteins were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting techniques. The chimera LFD1-PA4 and mixed LFD1+PA4 proteins were injected four times into mice and antibody production was relativity evaluated by enzyme-linked immunosorbent assay (ELISA) test. The results showed that both chimeric and mixed proteins are immunogenic, but LFD1-PA4 has a higher potential to stimulate mice immune system. LFD1-PA4 chimeric protein induced a higher immune response than LFD1+PA4 mixed protein and elicited antibody responses to LF and edema factor (EF), therefore, it holds promise to be a more effective trivalent vaccine candidate to use in anthrax prevention.

The immune response of aged broiler breeder hens is influenced by many factors including obesity and aged lymphatic organs, but may improve by increasing the bioavailability of various nutrients such as zinc (Zn). Dietary supplementation of phytase can improve Zn availability in senescent broiler breeder hens. The aim of this study was to investigate the effect of supplementary zinc oxide (ZnO) and phytase in a maize-soybean meal-based diet on immune responses of broiler breeder hens. In a 2 × 4 factorial arrangement, a total of 128 hens were randomly assigned into eight groups. The birds received two levels of phytase (0 or 300 U/kg diet) and four levels of ZnO (30, 60, 90, and 120 mg/kg diet) for 13 successive wk (59-72 wk of age). Results showed that phytase supplementation significantly increased immunoglobulin M (IgM), cutaneous basophil hypersensitivity (CBH) responses, total number of leukocytes, percentage of lymphocytes, and heterophil to lymphocyte ratios. The percentage of basophils and monocytes, however, decreased with phytase supplementation. Supplementation of ZnO increased anti-sheep red blood cells (SRBC) antibody titer, IgM, CBH responses, the total number of leukocytes, and the percentage of lymphocytes. Dietary supplementation of ZnO decreased the percentage of heterophil, and heterophil to lymphocyte ratio. A significant interaction effect of phytase and ZnO was found on the total number of leukocytes and percentage of lymphocytes. Dietary supplementation of ZnO (90 mg/kg diet) and phytase had some positive effects on improving immune responses in broiler breeder hens.

Sperm cryopreservation is an important tool for breed improvement, nonetheless, spermatozoids of rams are extremely sensitive to cryopreservation. The present research was to compare a liposome-based (OptiXcell: OX) diluent, a commercial TRIS-egg yolk (Optidyl: OP) and a citrate egg yolk-based (CY) diluent on ovine semen quality through the cryopreservation process. Semen was collected from four sexually mature Dorper rams during the natural breeding season. After collection, semen was evaluated and diluted in OX, OP or CY diluent and was cooled from 37°C to 4°C for 2 h (refrigerated semen, RS), after that semen-filled straws were placed in liquid nitrogen (LN) vapour for 10 min, then immersed into LN at -196°C (cryopreserved semen, CS) and stored until evaluation. For fresh semen (FS), similar values (P>0.05) were obtained from the 3 diluents [motility (4.2 ± 0.3), viability (75.4 ± 3.2), hypo-osmotic swelling test (HOST) (59.2 ± 2.1), and normality (84.7 ± 3.5)]. The motility values were higher for RS with OX and CY (4.0 ± 0.2 and 3.6 ± 0.3, respectively) compared to OP (3.0 ± 0.21; P<0.05). The viability was reduced after refrigeration and freezing (P<0.05). Refrigerated semen viability was similar for OX (65%), CY (63%) and OP diluents (60%; P>0.05), but for frozen semen, viability was lower in the CY diluent (P<0.05). Membrane integrity (HOST) in OX (53.6 ± 1.7) was similar to that in OP (50.7 ± 1.5; P>0.05) but higher than in CY (48.7 ± 1.5; P<0.05). No difference was found between the OX diluents and those made with egg yolk in terms of sperm parameters; however, the OX diluent was more efficient in protecting the integrity of membrane in freezing/thawing semen.

Orbifloxacin is being widely used in China to treat fish infections in an extra-label manner, which may cause its potential residues in edible tissues. The purpose of this study was to determine the disposition kinetics of orbifloxacin in crucian carp (Carassius auratus) following intramuscular administration for its further safe application in aquaculture industry. Tissue samples of skin, muscle, kidney, and liver were collected from six crucian carps reared at 25°C at 5, 10, 15, 30, 45 min, 1, 2, 4, 6, 12, 24, 48, 72, and 96 h following a single intramuscular injection at 7.5 mg/kg body weight (BW). The orbifloxacin concentrations in tissues were determined using a high-performance liquid chromatography (HPLC) method with a fluorescence detector, then average concentrations versus time data were subjected to non-compartmental analysis to obtain the kinetic parameters. The peak concentration of 5.68 ± 0.03 µg/g was calculated in kidney at 2 h, followed by muscle (5.51 ± 0.01 µg/g) at 4 h, liver (4.84 ± 0.20 µg/g) at 2 h, and skin (4.27 ± 0.08 µg/g) at 4 h. Area under concentration-time curve was calculated as 79.22, 94.72, 118.65, and 129.02 h·µg/g in kidney, liver, skin, and muscle, respectively. And the elimination half-lives were determined as 18.17, 18.41, 18.77, and 19.11 h in skin, kidney, muscle, and liver, respectively. It was shown that orbifloxacin was well distributed into tissues while relatively slowly eliminated in crucian carp reared at 25°C following a single intramuscular injection.

Orexin A, a small-molecule peptide, can regulate female hormones, but limited evidence for its mechanism of activity exists in ovine. The objective of this study was to investigate the effect of orexin A on progesterone (P4) secretion in cultured granulosa of sheep follicles. Sheep ovarian granulosa were isolated and identified, pre-incubated with luteinizing hormone (LH) (2.5 IU/ml), follicle-stimulating hormone (FSH) (2.5 IU/ml), or oestrogen (1 µg/ml); and cultured in vitro. The pretreated sheep ovarian granulosa were subsequently cultured with different concentrations (1 nM, 10 nM, 58 nM, 100 nM, and 145 nM) of orexin A for varying amounts of time (0 h, 24 h, 48 h, and 72 h). Then, the expression levels of P4, steroidogenic acute regulatory protein (StAR), 3β-Hydroxysteroid dehydrogenase (3β-HSD) and cytochrome P450 (CYP11) were determined. The results showed that the sheep ovarian granulosa were correctly identified. The different concentrations of orexin A promoted the secretion of P4 from granulosa in the ovine ovary compared with that in the control. The expression of StAR, 3β-HSD and P450 (CYP11) gradually increased, and then decreased with increasing concentrations of orexin A, but the expression of P450 (CYP11) decreased with the increase of time. These results revealed that orexin A promotes the secretion of P4 by regulating the expression of StAR, 3β-HSD, and P450 (CYP11). Understanding the mechanism underlying the promotion of P4 by orexin A could open new therapeutic possibilities in the treatment of hormone homeostasis.

Besnoitiosis caused by Besnoitia besnoiti is regarded as a re-emerging disease in cattle because of the increased number of cases and geographical distribution in many European countries. The present study was conducted to determine the presence of B. besnoiti in cattle in the Eastern and Southeastern Anatolia of Turkey. Blood samples were collected from 450 cattle in the provinces of Mus, Van, Siirt, and Diyarbakir. PrioCHECK®Besnoitia Ab 2.0 enzyme-linked immunosorbent assay (ELISA) kit was used to detect specific anti-B. besnoiti antibodies in the serum samples. ‏Twelve (2.7%) of the 450 asymptomatic cattle were seropositive against B. besnoiti. In cattle, the prevalence rates were 0%, 3.7%, 3.4%, and 1.1% in Mus, Siirt, Diyarbakir, and Van provinces (P>0.05), respectively. This study is the first to investigate the presence of B. besnoiti in cattle raised in the Eastern and Southeastern Anatolia of Turkey. Although the ELISA test revealed some positive cases, concrete evidence for the establishment of clinical B. besnoiti infection could not be verified. More comprehensive analysis would be necessary to determine the significance of the present observations.

The use of conventional stains including Giemsa, Wright and Leishman have become an essential tool for differential diagnosis of cells in peripheral blood. The aim of the study was to develop a new combination of Leishman-Giemsa (L&G) stain for avian blood smears and to compare its efficacy with conventional staining methods. Three sets of peripheral blood smears, one smear for L&G stain and two other smears for Leishman and Giemsa stains, created from 50 broiler chickens blood samples. All the three sets of slides were blind screened by two expert clinical pathologists and scored based on the staining characteristics (4 parameters) such as nuclear features of red blood cell (RBC) and white blood cell (WBC), cytoplasmic features and cytoplasmic granularity of WBC. The average grading score assigned by two experts for each staining method were compared. The average grading score of two conventional Leishman and Giemsa staining methods were significantly lower (P<0.001) than new L&G staining method in avian nuclear features of the RBC and WBC. The L&G stain gave a better clarity of nuclear features of avian RBC and WBC. The new L&G staining technique created significant differences (P<0.001) in cytoplasmic features of avian WBC compared to the other two methods. For the first time, the results of the present study showed that the avian blood cells are more desirable stained with a new combination of L&G stain. In addition, it gives a better nuclear and cytoplasmic differential staining than the conventional Giemsa and Leishman stains when used alone.

Autosomal-dominant polycystic kidney disease (ADPKD) is the most prevalent inherited genetic disease of cats, predominantly affecting Persians and Persian-related cats. The purpose of this study was to determine prevalence of polycystic kidney disease (PKD) in Persian cats in Iran, and also to assess the relationships between PKD and gender, age as well as clinical and paracilinical manifestations. Sonographic screening examination was performed on all healthy and unhealthy Persian and Persian-related cats referred to Small Animal Hospital of Faculty of Veterinary Medicine, University of Tehran, from April 2014 to May 2015. Cats were classified as positive when at least one anechoic cavity was found in at least one kidney. Of 76 Persian and Persian-related cats submitted for PKD ultrasound screening, 36.8% were found to have the disease and 63.2% were negative. Therefore, the prevalence of PKD was estimated 36.8% in Persian and Persian related cats in Tehran, Iran, which is approximately similar to prevalence in other parts of the world. Furthermore, there was a significant correlation between PKD and age, as in affected cats the detection probability of renal cysts in sonography was increased in older animals. For each year increase in age, the detection probability of PKD in sonography was increased about 2.62 times. The prevalence of the PKD amongst Persian cats in Iran is relatively high, and insufficient attention to incidence and prevalence of PKD especially in breeding programs, would spread the disease throughout in Persian cats.