Seeking a niche: putative contributions of the hfq and bacA gene products to the successful adaptation of the brucellae to their intracellular home

Long-term residence of the brucellae in the phagosomal compartment of host macrophages is essential to their ability to produce disease in both natural and experimental hosts. Correspondingly, the Brucella spp. appear to be well adapted to resist the multiple environmental stresses they encounter in their intracellular home. This brief review will focus on the contributions of the hfq and bacA gene products to this adaptation. Studies with Brucella hfq mutants suggest that stationary phase physiology is critical for successful long-term residence in host macrophages. Analysis of Brucella bacA mutants, on the other hand, reveal very striking parallels between the strategies employed by the rhizobia to establish and maintain protracted intracellular residence in their plant host and those used by the brucellae during their long-term survival in the phagosomal compartment of host macrophages.     

Brucellosis: a short review of the disease situation in Paraguay

A short review of the brucellosis situation, its control and eradication programs are presented. Data from over 1.2 milliom samples collected from more than 50,718 groups of cattle over a period of over 20 years (1979–2000) illustrates that over the last few years the number of individual reactors remain constant at around 3–4%. The percentage of reactive groups of animals decreased over these years, reflecting a better disease management and possibly an improved general education, handling of information on the immune (vaccination) status of animals and testing practices. Reported zoonotic cases are presented, as well as control and eradication programs, including utilization of vaccines.     

Bovine monocytes and a macrophage cell line differ in their ability to phagocytose and support the intracellular survival of Mycobacterium avium subsp. paratuberculosis

Bovine monocytes exhibited a greater ability to phagocytose Mycobacterium avium subsp. paratuberculosis (i.e. greater percentage of infected cells, and more bacilli per infected cell), than did a bovine macrophage cell line (BoMac). Phagocytosis of M. paratuberculosis by monocytes, but not the cell line, was significantly enhanced by the addition of autologous serum. Following ingestion, the numbers of viable M. paratuberculosis cells in monocytes increased during the first 4 days and then declined between day 4 and day 8 after infection, as determined by a radiometric method. In contrast, BoMac cells were not permissive for bacillary multiplication; the numbers of M. paratuberculosis remained largely unchanged in the cell line during the 8 day incubation period. The numbers of microscopically visible acid-fast bacilli increased with time in monocytes but not in the macrophage cell line. These observations suggest that replication and inactivation of bacilli may both occur in monocytes. The differing abilities of bovine monocytes and the macrophage cell line to ingest and restrain the intracellular growth of M. paratuberculosis provide contrasting model systems for investigating how M. paratuberculosis enters and persists within its preferred niche, the mononuclear phagocyte.     

Sheep grazing causes shift in sex ratio and cohort structure of Brandt's vole: Implication of their adaptation to food shortage

Livestock grazing has been demonstrated to affect the population abundance of small rodents in grasslands, but the causative mechanism of grazing on demographic parameters, particularly the age structure and sex ratio, is rarely investigated. In this study, we examined the effects of sheep grazing on the cohort structure and sex ratio of Brandt's vole (Lasiopodomys brandtii) in Inner Mongolia of China by using large manipulative experimental enclosures during 2010–2013. Our results indicated that sheep grazing significantly decreased the proportion of the spring‐born cohort, but increased the proportion of the summer‐born cohort. Grazing increased the proportion of males in both spring and summer cohorts. In addition, we found a negative relation between population density and the proportion of the overwinter cohort. Our results suggest that a shift in the cohort structure and the sex ratio may be an important strategy for small rodents to adapt to changes in food resources resulting from livestock grazing.     

Effects of yard weaning and training on the behavioural adaptation of cattle to a feedlot

Two experiments were conducted to determine the effect of weaning in small yards, with or without a feed bunk training procedure, on the subsequent behaviour and performance of Bos taurus steers in a feedlot. A reduction in the incidence of bovine respiratory disease (BRD) was the primary objective. In each experiment, about 200 male beef calves (Angus × Hereford and Hereford) were separated from their mothers at 7–9 months of age and allocated to one of three matched weaning treatment groups. The treatments were (1) yard weaning with hay or silage, (2) yard weaning with hay or silage plus a novel handling procedure to train the animals to be able to find a grain ration in a trough, and (3) paddock weaning without supplement or handling according to common industry practice in southeastern Australia. Paddock weaning is the practice of abrupt separation of cows and calves followed by return to separate pasture paddocks, whereas yard weaning involves abrupt separation but the calves remain in the yards for several days. Experimental vaccines against the major BRD pathogens were given to half of each group 1–2 months prior to entry into a commercial feedlot. The yard weaned and yard trained cattle had a significantly greater weight gain in the first month and over the 90-day feeding period than the paddock weaned control groups. There was no difference between the groups in pre-feedlot weight gain. The yard trained groups showed greater feeding activity during the first few days in the feedlot, but were not significantly different in weight gain from yard weaned. The vaccination treatment also significantly improved the weight gain in the first month and over 80 days. The combination of yard weaning and vaccination produced the highest weight gains overall. There was consistently less morbidity in the yard weaned groups compared to paddock weaned controls. The morbidity in yard trained groups was intermediate between these two. Weaning in small yards and the appropriate use of effective BRD vaccines 1–2 months before feedlot entry are recommended for B. taurus feeder steers in southeastern Australia to minimise sickness and improve productivity in the feedlot. Associated benefits are reduced risks of antibiotic residues and of animal welfare problems.     

Diagnosis of Canine Brucellosis: Comparison between Serological and Microbiological Tests and a PCR Based on Primers to 16S-23S rDNA Interspacer

A pair of primers directed to 16S-23S rDNA interspacer (ITS) was designed directed to Brucella genetic sequences in order to develop a polymerase chain reaction (PCR) putatively capable of amplifying DNA from any Brucella species. Nucleic acid extracts from whole-blood from naive dogs were spiked with decreasing amounts of Brucella canis RM6/66 DNA and the resulting solutions were tested by PCR. In addition, the ability of PCR to amplify Brucella spp. genetic sequences from naturally infected dogs was evaluated using 210 whole-blood samples of dogs from 19 kennels. The whole-blood samples collected were subjected to blood culture and PCR. Serodiagnosis was performed using the rapid slide agglutination test with and without 2-mercaptoethanol. The DNA from whole blood was extracted using proteinase-K, sodium dodecyl sulphate and cetyl trimethyl ammonium bromide followed by phenol–chloroform purification. The PCR was capable of detecting as little as 3.8 fg of Brucella DNA mixed with 450 ng of host DNA. Theoretically, 3.8 fg of Brucella DNA represents the total genomic mass of fewer than two bacterial cells. The PCR diagnostic sensitivity and specificity were 100%. From the results observed in the present study, we conclude that PCR could be used as confirmatory test for diagnosis of B. canis infection.     

Polymorphisms of a scrapie-associated fibril protein (PrP) gene and their association with susceptibility to experimentally induced scrapie in Cheviot sheep in the United States.

The duration of the incubation period for scrapie, a fatal transmissible neurodegenerative disorder of sheep and goats, is mainly determined by the Sip gene, which has 2 alleles (sA--susceptible and pA--resistant). A diagnostic test is not available to detect scrapie in live animals. We analyzed genomic DNA extracted from frozen sheep brains collected from Cheviot sheep of the United States that had been inoculated with the SSBP/1 scrapie inoculum. Digestion of the DNA with EcoRI or HindIII followed by the addition of a scrapie-associated fibril protein (PrP)-specific marker probe, yielded fragments of 6.8 (e1) and 4.0 (e3) kb, or 5.0 (h1) and 3.4 (h2) kb, respectively. Fragments e1 and h2 were associated with the histopathologic diagnosis of scrapie, and fragments e3 and h1 were associated with survival. A valine/alanine polymorphism within the PrP coding region that resulted in a BspHI site was further used to determine the genotype of these Cheviot sheep. Digestion of polymerase chain reaction fragments with BspHI resulted in an undigested fragment b- (0.840 kb), digested fragments b+ (0.460 and 0.380 kb), or both types of fragments. Survival time of b+/b+ homozygous sheep was significantly (P < 0.01) shorter (218 +/- 26.0 days) than survival time for b-/b- sheep (> 700 days after inoculation). Results indicated that b+ and b- are markers for the Sip sA and pA alleles, respectively. The intermediate duration of the incubation period for heterozygous sheep (b+/b-; 342.9 +/- 25.3 days) indicated that the Sip sA allele is expressed codominantly to the Sip pA allele.     

鸡舍环境耐药细菌气溶胶及其向环境传播的研究

通过对舍内粪便、空气和舍外环境(10、50、100m)中的空气分离到的细菌鉴定，包括细菌DNA分子生物学检测及药物敏感实验以及细菌含量统计学分析，证明三者之间存在着内在联系；舍内环境微生物气溶胶包括耐药细菌通过舍内外气体交换排往鸡舍周围环境，舍周边环境在一定范围内受到生物污染。研究显示，在粪便、舍内空气、舍外环境10、50、100m空气中分离的金黄色葡萄球菌和链球菌80％以上对氯霉素(CMP)和对苯唑青霉素(OXA)耐药；粪便和舍内外环境空气中的80％以上的大肠杆菌和沙门氏菌对氨苄青霉素(AMP）、氯霉素(CMP)、头孢唑啉(CFZ)均为抑制。证明该鸡舍曾经使用过这些药物治疗或作为添加剂在饲料中添加，鸡群产生了耐药细菌菌株，并向环境传播。测量数据统计结果指出，十里河鸡场鸡舍内环境空气中的大肠杆菌、沙门氏菌、金黄色葡萄球菌和链球菌与舍外环境中相应菌群含量比较差异显著或极显著(P＜0．05或P＜0．01)，但舍外10、50、100m之间的相应菌群比较差异不显著(P＞0．05)，这说明鸡舍内环境细菌气溶胶含量产生多，含量高，菌源强度大；另一方面，反应了鸡舍内环境微生物气溶胶包括耐药细菌能向舍外环境扩散，位舍外环境在一定的范围内受到动物源性生物污染，影响范围大于100m。通过对鸡舍环境微生物气溶胶来源的探讨发现，粪便中的大肠杆菌、沙门氏菌、金黄色葡萄球菌和链球菌与舍内空气相应菌群数量关系比较，呈现正相关(r=0.99)。这从一个侧面揭示了粪便可能是微生物气溶胶的主要来源。     

Determining the relationship between restorative potential and size of a gene bank to alleviate the risks inherent in a scrapie eradication breeding programme

Breeding plans are being adopted in many countries to eradicate scrapie from the sheep populations. The European plans are based on selection for increased frequency of the resistant ARR haplotype and removal of the most scrapie susceptible haplotype (VRQ). This approach of a genotype eradication breeding programme incurs a number of risks including those associated with novel spongiform encephalopathy causing agents that are as yet unknown, but may emerge in the future and the possible loss of favourable attributes of a breed either through linkage or pleiotropic effects when selecting on a specific haplotype. Semen gene banks have been proposed to provide the potential for future reintroduction of removed haplotypes. Computer simulations were run for hill, crossing and terminal breed types to investigate the restorative potential of a gene bank required for the reintroduction of an alternative haplotype in the future. Differences existed in the restorative potential between the different breed types considered. This was largely dictated by variable levels of fertility, litter size, survival and mating ratio. It was shown that terminal breeds would require two and a half times as many semen straws as hill breeds to produce the same frequency of ewes homozygous for the desired haplotype in the population after a ten year reintroduction period.     

基于小鼠表达谱芯片和比较基因组学分析猪生长发育相关候选基因集

为了在更宽、更深的层面上研究猪生长发育机制,得到以通路为单位的候选基因集以克服传统单基因分析方法的缺点。试验利用模式生物小鼠中的芯片技术,通过基因集富集分析方法(GSEA法)分析基因芯片,确定候选通路。对候选通路成员与猪生长发育相关的数量性状基因座(QTL)数据库进行映射,缩小其作用跨度区域,找出作用的关键基因集。结果表明:得到了56个与猪生长发育相关的候选基因集。     

New data about home range and movements of Oligoryzomys flavescens (Rodentia: Cricetidae) help to understand the spread and transmission of Andes virus that causes Hantavirus Pulmonary Syndrome

Hantavirus pulmonary syndrome is an emerging infectious disease caused by viruses of the genus Orthohantavirus. The rodent Oligoryzomys flavescens is distributed along four countries of South America. In Argentina, O. flavescens acts as a reservoir of three genotypes of ANDV orthohantavirus. The aims of this work were to estimate home range size and movements—with spool‐and‐line and radiotelemetry—of infected and non‐infected O. flavescens in order to understand the spread and transmission of the virus. O. flavescens use a wide area to satisfice its requirements, reaching a home range of 1.82 ha during spring. Orthohantavirus infection did not change the behaviour of individuals. We observed a great overlapping in the home range of infected and non‐infected individuals resulting in a high probability of virus dispersion on rodent population. These results show that human health risks could be high on island environments and knowledge about the movement ecology of O. flavescens provides useful information on prevention.     

Decision-making on the use of diverse combinations of agricultural products and natural plants in pig feed: a case study of native pig smallholder in northern Thailand

We have identified the feed offered to native pigs in a case study of smallholder in northern Thailand. We examined the types and fresh weights of pig feed over two 10-day periods in household A, in September 2006 (rainy season) and in December 2006 and January 2007 (dry season). The study results are as follows. (1) They offered 18 types of feed in total during the rainy and dry seasons, of which seven types were common to the rainy and dry seasons, five types were offered during the rainy season only, and six types during the dry season only. (2) They offered agricultural products as 34% of feed (rainy season) and 61% of feed (dry season), and natural plants used exclusively for pig feed as 66% of feed (rainy season) and 39% of feed (dry season). (3) The feed combinations at each feeding time differed 80% of the time during both the rainy and dry seasons. These results show not only that they offered diverse combinations of agricultural products and natural plants as pig feed, but also that they changed feed kinds in both the rainy and dry seasons.     

Blue sheep in the Annapurna Conservation Area,Nepal: habitat use,population biomass and their contribution to the carrying capacity of snow leopards

The Himalaya region of Nepal provides a habitat for the endangered snow leopard (Panthera uncia) and its principal prey species, the blue sheep (Pseudois nayaur). The aim of this study was to describe the habitat, the distribution and the population structure of blue sheep, and to estimate their contribution to the carrying capacity of snow leopard in the upper Mustang region of Nepal. Blue sheep were recorded at altitudes from 3209–5498 m on slopes with gradients of 16–60° and aspects of 40°NE to 140°SE. A total of 939 blue sheep were counted in the upper Mustang region, and 98 were counted in the Yak Kharka region of Manang district; however, upper Mustang had the lowest population density of blue sheep recorded within their distribution range in Nepal (0.86 blue sheep/km²). The results of the study show that a higher density of blue sheep is associated with greater plant species diversity. The most important species present in the blue sheep habitat were Kobresia pygmaea, Artemesia spp., Lonicera spp., Lancea tibetica, Poa spp., Astragalus spp. and Ephedra gerardiana. It is estimated that the existing blue sheep population biomass of approximately 38 925 kg in the upper Mustang region could support approximately 19 snow leopards (1.6 snow leopards/100 km²).     

Death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer and their progeny: a result of nationwide survey in Japan

To obtain the data concerning death losses due to stillbirth, neonatal death and diseases in cloned cattle derived from somatic cell nuclear transfer (SCNT) and their progeny produced by Japanese institutions, a nationwide survey was carried out in July-August, 2006. As a result, lifetime data concerning 482 SCNT cattle (97.5% of cattle produced in the country at that time) and 202 progeny of SCNT cattle were accumulated and the death loss of these cattle was analyzed. Although 1/3 of delivered SCNT calves died during the perinatal period due to stillbirth and neonatal death, incidence of death loss due to diseases in SCNT cattle surviving more than 200 days after birth seems to be the same as these in conventionally bred cattle. In contrast, progeny of SCNT cattle showed the same level in death loss as observed in conventionally bred cattle throughout their lifetime. These results suggest that robust health would be expected in SCNT cattle surviving to adulthood and their progeny.     

Use of the Capripoxvirus homologue of Vaccinia virus 30 kDa RNA polymerase subunit (RPO30) gene as a novel diagnostic and genotyping target: development of a classical PCR method to differentiate Goat poxvirus from Sheep poxvirus

Sheep poxvirus (SPPV), Goat poxvirus (GTPV) and Lumpy skin disease virus (LSDV) are Capripoxviruses (CaPVs) responsible for causing severe poxvirus disease in sheep, goats and cattle, respectively. Serological differentiation of CaPVs is not possible and strain identification has relied on the implicitly accepted hypothesis that the viruses show well defined host specificity. However, it is now known that cross infections can occur and authentication of identity based on the host animal species from which the strain was first isolated, is not valid and should be replaced with molecular techniques to allow unequivocal strain differentiation. To identify a diagnostic target for strain genotyping, the CaPV homologue of the Vaccinia virus E4L gene which encodes the 30 kDa DNA-dependent RNA polymerase subunit, RPO30 was analyzed. Forty-six isolates from different hosts and geographical origins were included. Most CaPVs fit into one of the three different groups according to their host origins: the SPPV, the GTPV and the LSDV group. A unique 21-nucleotide deletion was found in all SPPV isolates which was exploited to develop a RPO30-based classical PCR test to differentiate SPPV from GTPV that will allow rapid differential diagnosis of disease during CaPV outbreaks in small ruminants.