Pathogenesis of natural goat scrapie: modulation by host PRNP genotype and effect of co-existent conditions

After detection of a high prevalence of scrapie in a large dairy goat herd, 72 infected animals were examined by immunohistochemistry with prion protein (PrP) antibody Bar224 to study the pathogenesis of the infection. Tissues examined included the brain and thoracic spinal cord (TSC), a wide selection of lymphoreticular system (LRS) tissues, the distal ileum and its enteric nervous system (ENS), and other organs, including the mammary gland. The whole open reading frame of the PRNP gene was sequenced and antibodies to caprine arthritis-encephalitis virus (CAEV) infection were determined. Unexpectedly, accumulation of disease-associated PrP (PrP^d) in the brain was more frequent in methionine carriers at codon 142 (24/32, 75.0%) than amongst isoleucine homozygotes (14/40, 35.0%). The latter, however, showed significantly greater amounts of brain PrP^d than the former (average scores of 9.3 and 3.0, respectively). A significant proportion of the 38 goats that were positive in brain were negative in the ENS (44.7%) or in the TSC (39.5%). These results, together with the early and consistent involvement of the circumventricular organs and the hypothalamus, point towards a significant contribution of the haematogenous route in the process of neuroinvasion. Chronic enteritis was observed in 98 of the 200 goats examined, with no association with either scrapie infection or presence of PrP^d in the gut. Lymphoproliferative interstitial mastitis was observed in 13/31 CAEV-positive and scrapie-infected goats; PrP^d in the mammary gland was detected in five of those 13 goats, suggesting a possible contribution of CAEV infection in scrapie transmission via milk.     

State-of-the-art review of goat TSE in the European Union, with special emphasis on PRNP genetics and epidemiology

Scrapie is a fatal, neurodegenerative disease of sheep and goats. It is also the earliest known member in the family of diseases classified as transmissible spongiform encephalopathies (TSE) or prion diseases, which includes Creutzfeldt-Jakob disease in humans, bovine spongiform encephalopathy (BSE), and chronic wasting disease in cervids. The recent revelation of naturally occurring BSE in a goat has brought the issue of TSE in goats to the attention of the public. In contrast to scrapie, BSE presents a proven risk to humans. The risk of goat BSE, however, is difficult to evaluate, as our knowledge of TSE in goats is limited. Natural caprine scrapie has been discovered throughout Europe, with reported cases generally being greatest in countries with the highest goat populations. As with sheep scrapie, susceptibility and incubation period duration of goat scrapie are most likely controlled by the prion protein (PrP) gene (PRNP). Like the PRNP of sheep, the caprine PRNP shows significantly greater variability than that of cattle and humans. Although PRNP variability in goats differs from that observed in sheep, the two species share several identical alleles. Moreover, while the ARR allele associated with enhancing resistance in sheep is not present in the goat PRNP, there is evidence for the existence of other PrP variants related to resistance. This review presents the current knowledge of the epidemiology of caprine scrapie within the major European goat populations, and compiles the current data on genetic variability of PRNP.     

Protective effect of the AT137RQ and ARQK176 PrP alleles against classical scrapie in Sarda breed sheep

The susceptibility of sheep to scrapie is under the control of the host’s prion protein (PrP) gene and is also influenced by the strain of the agent. PrP polymorphisms at codons 136 (A/V), 154 (R/H) and 171 (Q/R/H) are the main determinants of susceptibility/resistance of sheep to classical scrapie. They are combined in four main variants of the wild-type ARQ allele: VRQ, AHQ, ARH and ARR. Breeding programmes have been undertaken on this basis in the European Union and the USA to increase the frequency of the resistant ARR allele in sheep populations. Herein, we report the results of a multi-flock study showing the protective effect of polymorphisms other than those at codons 136, 154 and 171 in Sarda breed sheep. All ARQ/ARQ affected sheep (n = 154) and 378 negative ARQ/ARQ controls from four scrapie outbreaks were submitted to sequencing of the PrP gene. The distribution of variations other than those at the standard three codons, between scrapie cases and negative controls, was statistically different in all flocks. In particular, the AT₁₃₇RQ and ARQK₁₇₆ alleles showed a clear protective effect. This is the first study demonstrating a protective influence of alleles other than ARR under field conditions. If further investigations in other sheep breeds and with other scrapie sources confirm these findings, the availability of various protective alleles in breeding programmes of sheep for scrapie resistance could be useful in breeds with a low frequency of the ARR allele and would allow maintaining a wider variability of the PrP gene.     

The role of mathematical modelling in understanding the epidemiology and control of sheep transmissible spongiform encephalopathies: a review

To deal with the incompleteness of observations and disentangle the complexities of transmission much use has been made of mathematical modelling when investigating the epidemiology of sheep transmissible spongiform encephalopathies (TSE) and, in particular, scrapie. Importantly, these modelling approaches allow the incidence of clinical disease to be related to the underlying prevalence of infection, thereby overcoming one of the major difficulties when studying these diseases. Models have been used to investigate the epidemiology of scrapie within individual flocks and at a regional level; to assess the efficacy of different control strategies, especially selective breeding programmes based on prion protein (PrP) genotype; to interpret the results of scrapie surveillance; and to inform the design of surveillance programmes. Furthermore, mathematical modelling has played an important role when assessing the risk to human health posed by the possible presence of bovine spongiform encephalopathy in sheep. Here, we review the various approaches that have been taken when developing and analysing mathematical models for the epidemiology and control of sheep TSE and assess their impact on our understanding of these diseases. We also identify areas that require further work, discuss future challenges and identify data gaps.     

Vaccination with a genotype 1 modified live vaccine against porcine reproductive and respiratory syndrome virus significantly reduces viremia,viral shedding and transmission of the virus in a quasi-natural experimental model

The present study assessed the efficacy of vaccination against genotype 1 porcine reproductive and respiratory syndrome virus (PRRSV) in terms of reduction of the transmission. Ninety-eight 3-week-old piglets were divided in two groups: V (n = 40) and NV (n = 58) that were housed separately. V animals were vaccinated with a commercial genotype 1 PRRSV vaccine while NV were kept as controls. On day 35 post-vaccination, 14 NV pigs were separated and inoculated intranasally with 2 ml of a heterologous genotype 1 PRRSV isolate (“seeder” pigs, SP). The other V and NV animals were distributed in groups of 5 pigs each. Two days later, one SP was introduced into each pen to expose V and NV to PRRSV. Sentinel pigs were allocated in adjacent pens. Follow-up was of 21 days. All NV (30/30) became viremic after contact with SP while only 53% of V pigs were detected so (21/40, p < 0.05). Vaccination shortened viremia (12.2 ± 4 versus 3.7 ± 3.4 days in NV and V pigs, respectively, p < 0.01). The 50% survival time for becoming infected (Kaplan–Meier) for V was 21 days (CI_95% = 14.1–27.9) compared to 7 days (CI_95% = 5.2–8.7) for NV animals (p < 0.01). These differences were reflected in the R value as well: 2.78 (CI_95% = 2.13–3.43) for NV and 0.53 (CI_95% = 0.19–0.76) for V pigs (p < 0.05). All sentinel pigs (10/10) in pens adjacent to NV + SP pens got infected compared to 1/4 sentinel pigs allocated contiguous to a V + SP pen. These data show that vaccination of piglets significantly decrease parameters related to PRRSV transmission.     

Simulation analysis of the effect of herd immunity and age structure on infection of a cattle herd with bluetongue viruses in Queensland, Australia

A state-transition model based on Leslie matrix formulation was used to investigate the effects of herd immunity and age structure on the infection of a simulated cattle herd with bluetongue viruses under Australian climatic conditions. Increasing duration of immunity decreased the prevalence of infection. A duration of immunity of 33 months was consistent with prevalence estimates made from previous serological studies of bluetongue virus. Herd prevalence displayed slowly dampening cyclical variation over time (most pronounced when a short duration of immunity was simulated). Increasing calving and mortality risk rates in the simulated herd increased prevalence, whereas increasing age at first calving decreased prevalence. Manipulation of calving rates had the greatest effect on the predicted prevalence of infection in the herd. Simulation of a number of herd-management scenarios suggested that management systems in which cattle are bred early and where high calving rates are achieved are likely to contribute to high levels of infection with bluetongue viruses. Results confirm the importance of management factors in influencing the prevalence of infectious diseases in animal populations.     

1株野鸭源鸡贫血病病毒的分离鉴定及其编码区基因序列分析

利用MDCC细胞系从东北地区采集的死亡野生鸟类样品中分离获得1株禽类病毒,对该病毒进行特异性PCR、特异性间接免疫荧光法等系统鉴定后,证实该分离株为鸡贫血病病毒(CAV),命名为WDNE110501株.利用PCR方法克隆出其编码区基因片段,测序结果表明,WDNE110501株的编码区全长为1 823 bp,无碱基缺失或插入.并将该基因序列和推导的氨基酸序列与国内外已发表的34个CAV株编码区基因进行同源性和亲缘关系的比对分析,同源性为96.1％～99.8％;氨基酸的同源性为89.8％～99.7％,与国内毒株harbin的差异最小,与国外最近的是美国毒株98D02152.序列比较表明CAV的3个编码基因VP1、VP2和VP3均有一定程度变异,以VP1变异性最大,且在不同毒株间的这3个阅读框的氨基酸序列变异是互不相关的.这是首次在野生鸟类中分离出CAV病毒,提示了我们野生鸟类在鸡传染性贫血病病毒传播和分布中可能起到一定作用.     

A field study on the effect of the dietary use of a clinoptilolite-rich tuff, alone or in combination with certain antimicrobials, on the health status and performance of weaned, growing and finishing pigs

This study was conducted to assess the effect of dietary use of a clinoptilolite-rich tuff (Cp) on health status and performance of weaned, growing and finishing pigs and its compatibility during simultaneous oral administration of antimicrobials (AM) such as enrofloxacin (E) or salinomycin (S). Weaners (720) were assigned in 2 experimental groups and 4 subgroups based on the inclusion or not of Cp and AM in their feed (subgroups: NC, ES, Cp, Cp+ES) in order to evaluate their health status, under PWDS prevailing herd conditions. A second part of the trial aimed to the evaluation of piglet performance under conditions with minimized PWDS herd risks. For this purpose, a second set of 264 weaners were assigned in 2 groups and 4 subgroups, in a respective manner. All piglets remained on-trial until slaughtering age; Cp was incorporated in their feed at a rate of 2% from the day of weaning until slaughtering. The health status evaluation consisted in monitoring piglets for adverse effects related to Cp consumption, average daily diarrhoea scoring during weaning and mortality rate calculations throughout. Performance evaluation included individual weighing at the end of weaning, growing and fattening periods and feed consumption assessments. Average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ration (FCR) on a pen basis were further calculated. Cp ingestion was well tolerated by the piglets. Simultaneous administration of Cp and AM in feed, resulted in less severe forms of PWDS, which had a shorter clinical course (P<0.05). Mortality decreased (P<0.05) during the weaning period due to AM administration. Concerning mean pig body weight at the end of each production phase, both Cp and AM had favorable effects (P<0.05). ADG estimated for the whole observation period was improved (P<0.05) by Cp-use along with AM. FCR improvements (P<0.05) were noticed during the different stages of growth due to AM or Cp administration, while Cp/AM interaction was noticed only at weaning (P<0.05).     

Role of pet dogs and cats in the transmission of helminthic zoonoses in Europe, with a focus on echinococcosis and toxocarosis

The close emotional tie between people and companion animals is a beneficial relation known as the human-animal bond. However, pet dogs and cats can play an important role in the transmission of helminthic zoonotic agents such as the tapeworms Echinococcus and the roundworms Toxocara which are directly transmitted from pets to the human environment without the involvement of vectors or intermediate hosts. In humans, echinococcosis has emerged in Europe and toxocarosis is still persisting in large endemic areas despite the availability of highly efficient anthelminthics for dogs and cats. Ecological changes significantly contributed to these trends: the high wild fox populations and the high density of freely roaming dogs and cats maintain a permanent infection pressure of these and other parasites. Further, the establishment of urban recreational environments closer to natural ecological systems boosted vole populations that represent urban reservoirs for zoonotic helminths. A good understanding of the parasites’ biology and epidemiology including the transmission to humans is required for planning and implementing effective prevention strategies. The continuous education of veterinarians and the information of the pet owners by providing uniform recommendations are of priority importance. A close collaboration between veterinary and public health professionals in a ‘One Health’ concept is required.     

Effects of a topically applied anaesthetic on the behaviour,pain sensitivity and weight gain of dairy calves following thermocautery disbudding with a local anaesthetic

ABSTRACT

Aims: To compare the effect of a topically applied anaesthetic to no pain relief or meloxicam on the behavioural responses, pain sensitivity and weight gain of calves following disbudding with or without sedation.

Methods: A total of 364, 2–6 week-old calves from three commercial farms were systematically allocated to one of six treatment groups. All calves received a cornual nerve block prior to disbudding, with half restrained in a crate and half sedated with xylazine. Within these groups one third received no further treatment (control), one third were treated with meloxicam >10?minutes prior to disbudding and one third received a topical anaesthetic applied to the horn bud wounds following disbudding. The frequency of ear flicks, head shakes, head scratches and pain sensitivity of the wound were recorded on up to eight occasions over 24 hours after disbudding. Calves were weighed before, and 7 and 28 days after, disbudding to determine average daily weight gain (ADG).

Results: Compared to calves in the crate-control group, all other groups had reduced ear flicks at all times following disbudding (p?<?0.01). Treatment with meloxicam and topical anaesthesia in addition to sedation reduced head scratches compared to calves in the crate-control group (p?≤?0.013). At 22 hours after disbudding head shakes were reduced in sedated calves treated with topical anaesthetic compared to calves in the crate-control group (p?<?0.001). Pain sensitivity was lower in all sedated calves than unsedated calves (p?<?0.001). The ADG between Days 0–7 was 0.14 (95% CI?=?0.015–0.274) kg/day greater in sedated calves treated with meloxicam than calves in the crate-control group (p?=?0.03), and the ADG between Days 0–28 tended to be 0.06 (95% CI=?0.01–0.13) kg/day greater in sedated calves treated with topical anaesthetic than calves in the crate-control group (p?=?0.09).

Conclusion and clinical relevance: Sedation of calves for disbudding reduced the pain experienced in the following 24 hours. There was a benefit to providing calves with topical anaesthetic following disbudding on behavioural responses and pain sensitivity, which was similar to that of treating calves with meloxicam.     

Epistatic effects between pairs of the growth hormone secretagogue receptor 1a,growth hormone,growth hormone receptor,non‐SMC condensin I complex,subunit G and stearoyl‐CoA desaturase genes on carcass,price‐related and fatty acid composition traits in Japanese Black cattle

Growth hormone secretagogue receptor 1a (GHSR1a), growth hormone (GH), growth hormone receptor (GHR), non‐SMC condensin I complex, subunit G (NCAPG) and stearoyl‐CoA desaturase (SCD), are known to play important roles in growth and lipid metabolisms. Single and epistatic effects of the five genes on carcass, price‐related and fatty acid (FA) composition traits were analyzed in a commercial Japanese Black cattle population of Ibaraki Prefecture. A total of 650 steers and 116 heifers for carcass and price‐related traits, and 158 steers for FA composition traits were used in this study. Epistatic effects between pairs of the five genes were found in several traits. Alleles showing strain‐specific differences in the five genes had significant single and epistatic effects in some traits. The data suggest that a TG‐repeat polymorphism of the GHSR1a.5′UTR‐(TG)_n locus plays a central role in gene–gene epistatic interaction of FA composition traits in the adipose tissue of Japanese Black cattle.     

Effects of dietary supplementation with a chlorella by‐product on the growth performance,immune response,intestinal microflora and intestinal mucosal morphology in broiler chickens

This study aimed to determine the effect of different dietary levels of a Chlorella by‐product (CBP) on the growth performance, immune response, intestinal microflora and intestinal mucosal morphology of broilers. In total, 480 one‐day‐old broiler chickens were randomly allotted to four dietary treatments with four replicated pens consisting of 30 chicks. The basal diet was formulated to be adequate in energy and nutrients. Three additional diets were prepared by supplementing 25, 50 or 75 g/kg of CBP to the basal diet. The diets were fed to the broilers ad libitum for 35 days. Result indicated that increasing inclusion level of CBP improved BW gain (linear, p < 0.05). There was no effect of inclusion level of CBP in diets on total cholesterol, triglyceride, aspartate aminotransferase and alanine aminotransferase levels during the 35 days. Plasma IgG, IgM and IgA concentrations increased (linear, p < 0.05) with inclusion level of CBP in diets. Supplementation of CBP in the diets increased (linear, p < 0.05) the concentrations of Lactobacillus in the caecal content and decreased (linear, p < 0.05) the concentrations of Escherichia coli and Salmonella in the caecal content. Villus height increased (linear and quadratic, p < 0.05) with inclusion level of CBP in diets. Crypt depth increased (quadratic, p < 0.05) with inclusion level of CBP, and a decreased villus height: crypt depth ratio (quadratic, p < 0.05) was observed as inclusion level of CBP in diets increased. The results of the current experiment indicate that dietary supplementation of CBP improves growth performance of birds. Dietary CBP has improving Lactobacillus spp. concentrations in the gastrointestinal tract, plasma immunoglobulin concentrations and intestinal mucosal morphology.     

Comparison of the effects of topical administration of a fixed combination of dorzolamide-timolol to monotherapy with timolol or dorzolamide on IOP, pupil size, and heart rate in glaucomatous dogs

OBJECTIVE: To determine whether the combination multiple-dose dorzolamide-timolol administered topically has any greater effects on the reduction of intraocular pressure, pupil size, and heart rate in dogs with glaucoma than do either timolol or dorzolamide alone. PROCEDURE: Applanation tonometry, pupil size, and heart rate measurements were made at 7 a.m., 1 p.m., and 7 p.m. daily of 12 laboratory Beagles with inherited primary open-angle glaucoma during each active phase of this study. Timolol 0.5% was administered first twice daily for 4 consecutive days. Dorzolamide 2.0% was administered next three times daily for 4 consecutive days. The fixed combination of the two (timolol 0.5% and dorzolamide 2.0%) was administered twice daily for 4 consecutive days during the final week of the study. Between administration of each drug, a withdrawal period of at least 10 days was instituted. Statistical comparisons between the effects of the three drugs were performed. RESULTS: Intraocular pressure (IOP) was decreased with the administration of all three drugs: timolol alone, dorzolamide alone, and the combination of the two decreased IOP after 1 day of treatment 2.83 +/- 0.70 mmHg, 6.47 +/- 0.32 mmHg, and 6.56 +/- 0.37 mmHg, respectively. After 4 days of treatment, the IOP decreased even further: timolol alone, dorzolamide alone, and the combination of the two decreased IOP 3.75 +/- 0.88 mmHg, 7.50 +/- 0.29 mmHg, and 8.42 +/- 0.58 mmHg, respectively. Heart rate was significantly decreased with timolol (-11.9 +/- 2.0 bpm) and the combination preparation (-8.6 + 2.4 bpm), but not with dorzolamide (-3.7 +/- 1.8 bpm) alone. Pupil size was significantly decreased with timolol (-1.42 + 0.40 mm) and the combination preparation (-1.3 + 0.33 mm), but not with dorzolamide (0.97 +/- 0.36 mm) alone. CONCLUSIONS: The combination dorzolamide-timolol appears to be more effective at reducing intraocular pressure in glaucomatous dogs than is either timolol or dorzolamide alone.     

Effect of grazing pasture with a low selenium content on the concentrations of triiodothyronine and thyroxine in serum,and GSH-Px activity in erythrocytes in cows in Chile

AIM: To determine the effect of grazing pasture that had a low selenium (Se) concentration on serum concentrations of tri iodothyronine (T₃) and thyroxine (T₄), and erythrocyte glutathione peroxidase (GSH-Px) activity in dairy cows.

METHODS: Forty pregnant Friesian cows were grazed on pasture that contained 0.03–0.04 ppm Se on a dry matter (DM) basis. Two months before parturition, 20 cows were randomly selected and treated with 1 mg Se/kg bodyweight subcutaneously, as barium selenate (Group Se-S). The other group (Se-D) was not supplemented. Blood samples were taken before supplementation (-60 days) and 30, 60, 90, 180 and 270 days after parturition, for determination of concentrations of T₃ and T₄ in serum, and GSH-Px activity in erythrocytes.

RESULTS: Erythrocyte GSH-Px activity in the Se-D group was >60 U/g haemoglobin (Hb) throughout the experiment. Supplementation increased (p>0.05) activities to >130 U/g Hb throughout lactation. Mean serum concentrations of T₄ in Se-D and Se-S cows increased from 23.7 (SEM 0.7) and 23.4 (SEM 0.8) nmol/L, respectively, in the prepartum period to 69.6 (SEM 0.1) and 67.6 (SEM 0.2) nmol/L, respectively, at 180 days of lactation (p>0.01), and no effect of Se supplementation was evident. Serum concentrations of T₃ in Se-D cows decreased (p>0.05) from 1.6 (SEM 0.1) nmol/L prepartum to 1.0 (SEM 0.2) nmol/L at the beginning of lactation, and remained lower (p>0.05) than those in the Se-S cows which did not decrease after calving and ranged from 1.9 (SEM 0.1) to 2.4 (SEM 0.2) nmol/L throughout lactation.

CONCLUSIONS: Serum T₃ concentrations decreased during early lactation in unsupplemented cows grazing pastures low in Se (0.03–0.04 ppm) and both serum T₃ and erythrocyte GSH-Px activities were consistently lower throughout lactation compared with Se-supplemented cows. Se supplementation had no effect on serum T₄ concentrations.     

Effects of nitrate addition to a diet on fermentation and microbial populations in the rumen of goats,with special reference to Selenomonas ruminantium having the ability to reduce nitrate and nitrite

This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real‐time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real‐time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate‐ and nitrite‐reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen.     

The Influence of the Halothane Test on Heart Parameters,Oct-Activity,Acid-Base Balance and Blood Electrolytes in Halothane-Sensitive Pigs and in Pigs Pre Medicated with a β-Blocker (Propranolol)

The heart and liver functions, blood electrolytes and acid-base balance were studies in halothane-sensitive and nonsensitive pigs. An inhibition of the hyperthermia reaction was attempted by an i.v. injection of propranolol. The pulse rate of sensitive pigs was shown to be elevated already at the beginning of halothane narcosis. No other differences between sensitive and nonsensitive pigs were observed from the electrocardiogram. Ectopic beats and arythmia appeared only in 1 halothane-sensitive pig which died soon after the test. The serum OCT-activity showed no abnormalities. Hyperkalemia and acidosis occurred but the Ga⁺⁺ was not elevated in halothane-sensitive animals. The slight elevation in Na⁺ was thought to be caused by hemoconcentration. A propranolol injection only delayed but did not inhibit the halothane reaction.