Evaluation of the safety,immunogenicity and efficacy of three capripoxvirus vaccine strains against lumpy skin disease virus

The safety, immunogenicity and efficacy of three commercially available vaccines against lumpy skin disease (LSD) in cattle have been evaluated using a combination of vaccine challenge experiments and the monitoring of immune responses in vaccinated animals in the field. The three vaccines evaluated in the study included two locally produced (Ethiopian) vaccines (lumpy skin disease virus (LSDV) Neethling and Kenyan sheep and goat pox (KSGP) O-180 strain vaccines) and a Gorgan goat pox (GTP) vaccine manufactured by Jordan Bio-Industries Centre (JOVAC). The latter vaccine was evaluated for the first time in cattle against LSDV. The Ethiopian Neethling and KSGPO-180 vaccines failed to provide protection in cattle against LSDV, whereas the Gorgan GTP vaccine protected all the vaccinated calves from clinical signs of LSD. There was no significant difference in protective efficacy detected between two dosage levels (P = 0.2, P = 0.25, and P = 0.1 for KSGP, Neethling and Gorgan vaccines, respectively). Additionally, the Gorgan GTP vaccinated cattle showed stronger levels of cellular immune responses measured using Delayed-Type Hypersensitivity (DTH) reactions at the vaccination site indicating higher levels of immunogenicity produced by the GTPV vaccine in cattle, as opposed to the other two vaccines. This study indicated, for the first time, that the Gorgan GTP vaccine can effectively protect cattle against LSDV and that the Neethling and KSGP O-180 vaccine were not protective. The results emphasise the need for molecular characterization of the Neethling and KSGP O-180 vaccine seed viruses used for vaccine production in Ethiopia. In addition, the potency and efficacy testing process of the Ethiopian LSD Neethling and KSGP O-180 vaccines should be re-evaluated.     

Prevalence and Intensity of Paramphistomum Spp. In Cattle from South-Eastern Iran

Background:

Parasitological investigations on paramphistomosis were carried out over a 12-month period in the southeast of Iran to determine the prevalence and intensity of this disease.

Methods:

A total of 1000 cattle, Sistani breed (n= 450) and Brahman breed (n= 550) of all sex and age groups were inspected at random for the presence of paramphistomidae flukes in Zabol slaughterhouse from December 2012 to October 2013.

Results:

Paramphistomes were found in 369 of 1000 necropsied cows (36.9%; 95% CI: 30.1–41.9%), with significant higher prevalence of infection in Brahman breed than in Sistani breed (51% vs 19.3%). No significant correlation between prevalence, intensity of infection, sex and age of cattle was noted. Despite the difference in the seasonal variations of prevalence, and the relation between the intensity of infection and season, these were not statistically significant. The mean intensity of infection in Brahman breed was higher (652.66 ± 281.5) than Sistani breed (123.32 ± 32.2). The identification of stained trematodes to the species revealed 40, 20, 20, 15 and 5% Gastrothylax crumenifer, Cotylophoron cotylophorom, Paramphistomum cervi, Carmyerius spatiosus, Explanatum explanatum, respectively.

Conclusion:

The present results will contribute to our understanding of the epidemiology of paramphistomumosis in southeastern Iran.     

Development of an Indirect ELISA Using Different Fragments of Recombinant Ncgra7 for Detection of Neospora caninum Infection in Cattle and Water Buffalo

Background:

Dense granules are immunodominant proteins for the standardization of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagnosis of Neospora caninum in cattle and water buffalo.

Methods:

NcGRA7, from N. caninum tachyzoites was amplified. PCR product and pMAL-c2X plasmid were digested with EcoR1 restriction enzyme and expressed in Escherichia coli to evaluate its competence for detection of anti- N. caninum antibodies with ELISA in comparison with commercial IDEXX ELISA. Furthermore, 230 sera of presumably healthy cattle and water buffaloes (108 cattle and 122 water buffaloes) were analyzed by both tests to determine the agreement of these two procedures.

Results:

Sensitivities and specificities of NcGRA7-based ELISA were 94.64% and 90.38% respectively using sera of cattle, but were 98.57% and 86.54% in the case of buffaloes respectively. A good correlation between the results of IDEXX ELISA and ELISA based on recombinant NcGRA7 for detecting N. caninum antibodies was appeared. Analyzing by Mc Nemar′s showed that NcGRA7-based ELISA has acceptable capability to differentiate the positive results in comparison with IDEXX ELISA.

Conclusion:

NcGRA7-based ELISA considering utilized new fragment of genomic DNA is a good tool for serodiagnosis of anti- N. caninum antibodies for screening and epidemiological purposes on cattle herd and water buffaloes as well.     

Heavy Metal Residues in the Milk of Cattle and Goats During Winter Season

Cadmium (Cd), chromium (Cr), nickel (Ni) and lead (Pb) residues (mg/L) were determined in the milk of cattle and goats. The milk samples of cattle from area 1 have higher levels of residues than area 2; Cd 0.089 ± 0.002 vs. 0.062 ± 0.01 Cr 1.14 ± 0.046 vs. 0.995 ± 0.017 Ni 23.38 ± 0.564 vs. 21.407 ± 0.275 Pb 21.781 ± 0.172 vs. 15.958 ± 1.00. The residual levels of Cd (0.084 ± 0.003) and Pb (42.687 ± 0.051) have been found higher in goat milk. The Ni residues in cattle milk (22.395 ± 0.988) are higher than in goat milk (19.522 ± 0.011) while residues of Cr are non significantly different in both species.     

改良奶牛与传统蒙古黄牛的牛奶中雌激素和孕激素含量的比较

目的 测定我国目前市场上销售的改良奶牛的牛奶中雌激素和孕激素的含量,比较改良奶牛和传统蒙古黄牛所产牛奶的雌激素和孕激素含量.方法 用酶联免疫试剂盒测定牛奶中的雌酮、雌二醇、孕酮含量;并通过文献阅读和现场调查比较家庭散养的蒙古黄牛和饲养基地的改良奶牛在饲养和牛奶生产上的差异.结果 本研究中测定的蒙古黄牛奶中雌酮平均浓度为(98.5±12.4)pg/ml,雌二醇平均浓度为(24.6±3.0)pg/ml,孕酮平均浓度为(0.2±0.3)ng/ml.3种晶牌的市售奶之间,雌激素和孕激素的浓度高低不等,雌酮浓度分别为(150.2±8.4)、(131.3±16.3)、(128.9±13.0)pg/ml;雌二醇浓度分别为(35.4±2.2)、(30.3±3.1)、(30.0±2.0)pg/ml;孕酮浓度分别为(20.2±1.5)、(18.1±2.2)、(16.5±2.4)ng/ml.3种市场奶中的雌酮、雌二醇以及孕酮的含量均高于黄牛奶中的含量(雌酮此较:t=5.43,19.23,5.89;雌二醇比较:t=4.14,4.93,14.03;孕酮比较:t=28.47,32.73,22.82;P值均<0.05).文献阅读和现场调查发现家庭散养的传统黄牛在妊娠中后期不产奶,而奶牛饲养基地的改良奶牛在雌激素和孕激素水平显著升高的妊娠中后期继续产奶,改良奶牛的泌乳期明显延长,产奶量明显增加.结论 目前我国市场上销售的牛奶中含有一定量的雌激素和孕激素,含量高于蒙古黄牛生产的牛奶.这一结论不能否定牛奶的营养价值,旨在提醒大家关注牛奶中的雌激素和孕激素可能对健康产生的影响.     

The mycobacterial glycolipid glucose monomycolate induces a memory T cell response comparable to a model protein antigen and no B cell response upon experimental vaccination of cattle

Glycolipids are presented to T cells by human group 1 CD1 proteins, but are not used as subunit vaccines yet. Experimental immunizations with pure mycobacterial glucose monomycolate (GMM) and keyhole limpet haemocyanin (KLH) in cattle, a species which, unlike mice, expresses group 1 CD1, showed that GMM was equally efficient as KLH in generating T cell responses in blood, but not in the draining lymph node. Also, KLH induced strong antibody responses whereas GMM did not. These data suggest that non-overlapping T cell populations are targeted and demonstrate the potential of glycolipids as a special class of subunit vaccine candidates.     

Immunity of cattle following vaccination with a Mannheimia haemolytica chimeric PlpE-LKT (SAC89) protein

We developed several chimeric PlpE-leukotoxin (LKT) constructs containing the major epitope of Mannheimia haemolytica outer membrane lipoprotein PlpE (epitope R2) and the neutralizing epitope of M. haemolytica LKT (NLKT) [Ayalew et al. Mannheimia haemolytica chimeric protein vaccine composed of the major surface-exposed epitope of outer membrane lipoprotein PlpE and the neutralizing epitope of leukotoxin. Vaccine 2008;26(38):4955-61]. Vaccination of mice with these PlpE-LKT chimeric proteins stimulated anti-PlpE antibodies that caused complement-mediated bacteriolysis of M. haemolytica as well as neutralizing anti-LKT antibodies. Chimeric protein SAC89, which contains two copies of R2 and two copies of NLKT, generally stimulated the best overall responses in mice. The objectives of the current study were: (1) to determine through a dose titration study if vaccination of cattle with SAC89 stimulated antibodies to both PlpE and LKT and (2) evaluate SAC89-induced immunity against experimental M. haemolytica challenge of cattle. In the dose titration study, vaccine doses ranged from 100 to 400mug. SAC89 significant anti-M. haemolytica surface and LKT antibodies were detected following vaccination with each dose. The vaccination/challenge study was conducted with 30 weaned beef cattle distributed among four groups: Control (no vaccine), 100mug SAC89, M. haemolytica Bacterin, and SAC89+M. haemolytica bacterin. On day 42 after two vaccinations, cattle were challenged transthoracically with M. haemolytica. There was significant reduction (p<0.05) in lesion scores for the SAC89+bacterin-vaccinated group (74.6% reduction compared to control lesion scores) when compared to the other groups (34.7% and 35.6% reduction compared to control lesion scores). Evaluation of antibody responses demonstrated that the control group failed to develop antibody responses to M. haemolytica surface antigens or to LKT. Bacterin-vaccinated cattle developed anti-M. haemolytica antibodies after the second vaccination. SAC89- and SAC89+bacterin-vaccinated groups developed significant antibody responses 14 days after the first vaccination and further significant increases in antibodies after the second vaccination. CONCLUSIONS: Vaccination with the chimeric protein SAC89 in conjunction with a M. haemolytica bacterin stimulated significant protection against a severe transthoracic challenge with the bacterium.     

The influence of polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite—dichlorodiphenyldichloroethylene (DDE) on mRNA expression for NP-I/OT and PGA, involved in oxytocin synthesis in bovine granulosa and luteal cells

The effect of polychlorinated biphenyls (PCBs) congeners (PCB 77, PCB 126, PCB 153) and their technical mixture—Aroclor (Ar) 1248, as well as dichlorodiphenyltrichloroethane (DDT) and its metabolite—dichlorodiphenyldichloroethylene (DDE; two individual isomers p,p′- and o,p′- or their mixture, 95% and 5%, respectively) at the dose of 10 ng/ml each, on the gene expression of (a) oxytocin (OT) precursor—neurophysin–oxytocin (NP-I/OT) and (b) peptidyl glycine-α-amidating mono-oxygenase (PGA), the terminal enzyme in the pathway of OT synthesis, was studied. Granulosa cells from follicles >1 cm in diameter, collected on days 19–21 of estrous cycle, and luteal cells from corpora lutea (CL) collected on days 8–12 of the estrous cycle were used. The cells were incubated (6 h) with these xenobiotics and the expression of NP-I/OT and PGA genes was determined. All PCBs increased (P < 0.05) NP-I/OT gene expression in granulosa cells. Similarly, all PCBs but PCB 126 increased (P < 0.05) PGA gene expression in these cells. DDT and DDE increased (P < 0.05) gene expression of NP-I/OT in granulosa cells, while gene expression of PGA in these cells was stimulated (P < 0.05) by DDE only. The mRNA expression for NP-I/OT and PGA in luteal cells was increased (P < 0.05) by PCB 77 and PCB 153. Both DDE isomers and mixture also stimulated (P < 0.05) of NP-I/OT mRNA expression, while increase (P < 0.05) of PGA mRNA expression was elicited by incubation of these cells with DDE mixture and Ar 1248.Obtained data suggest that PCBs, DDT and DDE can affect the mRNA expression for NP-I/OT and PGA in bovine granulosa and luteal cells.     

Zinc and selenium status in cows with dermatophytosis

Dermatophytosis (ringworm), an infection of the superficial keratinized structures of the skin and hair, is the most common contagious skin disease in cattle. The infectious diseases caused by dermatophytes are mainly related to the enzymes product by these fungi. Conversely, elements such as zinc and selenium are involved in the regulation of immune responses to infection. There are rare reports about the possible role of zinc and selenium concentration in the pathogenesis of cattle dermatophytosis. Thus, this study was conducted in a humid area of Iran on 35 healthy and 35 infected cows. After diagnosis confirmation by direct microscopic examination and fungi isolation via inoculation on Sabouraud dextrose agar using skin scrap and broken hair samples of infected cows, the zinc and selenium concentration of serum and hair in both groups were determined by potentiometric stripping analyzer and atomic absorption spectrometry, respectively. Results showed that serum concentration of selenium and zinc in cattle with dermatophytosis were significantly lower (P < 0.05) than the healthy ones. Although hair concentration of selenium and zinc in infected cattle were lower than the healthy ones, the differences were not significant (P > 0.05). In conclusion, it seems that zinc and selenium have a determinant role in immune status and the response of animal’s immunity system to dermatophytosis.     

脑苷肌肽在新生儿缺氧缺血性脑病中的应用

目的探讨脑苷肌肽对新生儿缺氧缺血性脑病（HIE）的治疗作用。方法将HIE患儿随机分成治疗组和对照组。对照组36例给予常规治疗,治疗组34例在常规治疗基础上给予脑苷肌肽2ml/d静脉滴注,1次/d,观察临床表现和头颅CT恢复情况。结果治疗组临床症状恢复时间均较对照组明显缩短,差异有统计学意义（P〈0.05）;治疗组总有效率为90.0%,对照组为67.8%,差异有统计学意义（P〈0.05）;1～2周后复查头颅CT,治疗组脑组织低密度影全部消失,对照组有8例改善不明显,两组比较差异有统计学意义（P〈0.05）。结论脑苷肌肽对HIE有较显著的治疗作用。