Suspected primary lactose intolerance in neonatal foals

The intestine of neonatal mammals must be able to secrete lactase enzyme to hydrolyse lactose in its mother's milk. Failure to secrete lactase results in failure to digest lactose. This results in osmotic diarrhoea, weight loss, failure to thrive, lethargy, colic and abdominal distension. Lactose intolerance may occur secondarily to intestinal bacterial infection. Primary lactose intolerance is recognised in human infants. Here we report 2 cases of suspected primary lactose intolerance in foals. The foals responded well to oral administration of exogenous lactase enzyme.     

Prophylaxis using paromomycin of natural cryptosporidial infection in neonatal kids

The chemoprophylactic effects of paromomycin sulfate against natural cryptosporidiosis in young kids were investigated. Two studies were carried out using two groups of 18 and 12 animals in two pens. In each pen, kids were allocated to treated or control groups. The treatment consisted of oral paromomycin given at 100 mg kg⁻¹ body weight day⁻¹ for 11 consecutive days from 2 days of age. All kids were weighed at 2, 6 and 10 days of age. Infection was monitored by collecting fecal samples and staining fecal smears every 3–4 days from days 4 or 5 to days 15 or 19. The results clearly showed the efficacy of paromomycin in reducing cryptosporidial oocyst output. Moreover, paromomycin prevented clinical signs and mortality.     

Cryptosporidium parvum infection and management-based risk factors of dairy calves in Taiwan

Cryptosporidiosis is one of the major causes of diarrhea in calves. Cryptosporidium parvum is considered the most important calf diarrhea pathogen in the Cryptosporidium species. Not only could infected calves spread C. parvum, but infected adult cattle could also shed oocysts. The objectives of this study were (1) to investigate the prevalence of C. parvum in dairy herds in Taiwan, including calves, the dams in delivery enclosure, the floor, and the drinking water; (2) to clarify the relationship of diarrhea, management, and C. parvum infection. Twenty dairy herds in Taiwan were selected by random sampling, including 226 calves and 198 dams, and other environmental samples were collected. A questionnaire was filled out by the farm owners to collect information regarding the management of calves and the delivery enclosure. Hierarchical logistic regression was used to analyze the risk factors for C. parvum infection. The prevalence of C. parvum infection in calves was 26.5% (60/226), while in dams, it was 19.7% (39/198). The C. parvum infection in calves increased with environmental contamination of C. parvum and clinical signs of diarrhea, while it decreased with a yard provided in the delivery enclosure. In conclusion, the management of the delivery enclosure appears to be more important for preventing cryptosporidiosis in calves in Taiwan.     

Association of adrenocorticotrophin and cortisol concentrations with peripheral blood leukocyte cytokine gene expression in septic and nonseptic neonatal foals

Background

The hypothalamic‐pituitary‐adrenal (HPA) is influenced by the proinflammatory cytokines IL‐6, IL‐1β, and TNF‐α in critically ill humans. Information about the association of cytokines with the HPA axis in neonatal foals is lacking.

Hypothesis/Objectives

The objectives were to describe for hospitalized septic and nonseptic foals (1) temporal changes in blood concentrations of ACTH, and cortisol, and leukocyte cytokine gene expression, and (2) coassociation of these HPA axis hormones with blood leukocyte cytokine gene expression.

Animals

Hospitalized septic foals (N = 15) and hospitalized nonseptic foals (N = 11).

Methods

Blood samples, obtained from study foals at admission (T = 0), and 24 (T = 1), 48 (T = 2), 72 (T = 3), and 96 (T = 4) hours after admission, were processed to isolate RNA from leukocytes and to harvest plasma and serum for hormone assays. Plasma ACTH and serum cortisol concentrations were determined by radioimmunoassay. Leukocyte mRNA expression of IL‐1β IL‐6, IL‐8, IL‐10, and TNF‐α was determined using RT‐PCR.

Results

Cortisol concentrations were greater (P < .05) in foals at admission than at other time points. The expressions of IL‐8 and IL‐10 mRNA were lower (P < .05) at each time point in septic than in nonseptic foals. Among septic foals, ACTH was positively associated (P = .0026) with IL‐6 mRNA expression.

Conclusions

Sepsis influences secretion of the HPA axis hormones and expression of cytokines in foals. A positive association with the HPA axis and IL‐6 expression was detected. The clinical importance of these findings requires additional study.     

Equine respiratory viruses in foals in New Zealand

AIMS: To identify the respiratory viruses that are present among foals in New Zealand and to establish the age at which foals first become infected with these viruses. METHODS: Foals were recruited to the study in October/ November 1995 at the age of 1 month (Group A) or in March/ April 1996 at the age of 4-6 months (Groups B and C). Nasal swabs and blood samples were collected at monthly intervals. Nasal swabs and peripheral blood leucocytes (PBL) harvested from heparinised blood samples were used for virus isolation; serum harvested from whole-blood samples was used for serological testing for the presence of antibodies against equine herpesvirus (EHV)-1 or -4, equine rhinitis-A virus (ERAV), equine rhinitis-B virus (ERBV), equine adenovirus 1 (EAdV-1), equine arteritis virus (EAV), reovirus 3 and parainfluenza virus type 3 (PIV3). Twelve foals were sampled until December 1996; the remaining 19 foals were lost from the study at various times prior to this date. RESULTS: The only viruses isolated were EHV-2 and EHV-5. EHV-2 was isolated from 155/157 PBL samples collected during the period of study and from 40/172 nasal swabs collected from 18 foals. All isolations from nasal swabs, except one, were made over a period of 2-4 months from January to April (Group A), March to April (Group B) or May to July (Group C). EHV-5 was isolated from either PBL, nasal swabs, or both, from 15 foals on 32 occasions. All foals were positive for antibodies to EHV-1 or EHV-4, as tested by serum neutralisation (SN), on at least one sampling occasion and all but one were positive for EHV-1 antibodies measured by enzyme-linked immunosorbent assay (ELISA) on at least one sampling occasion. Recent EHV-1 infection was evident at least once during the period of study in 18/23 (78%) foals for which at least two samples were collected. SN antibodies to ERBV were evident in 19/23 (83%) foals on at least one sampling occasion and 15/23 foals showed evidence of seroconversion to ERBV. Antibodies to ERAV were only detected in serum samples collected from foals in Group A and probably represented maternally-derived antibodies. Haemagglutination inhibition (HI) antibody titres 1:10 to EAdV-1were evident in 21/23 (91%) foals on at least one sampling occasion and 16/23 foals showed serological evidence of recent EAdV-1 infection. None of the 67 serum samples tested were positive for antibodies to EAV, reovirus 3 or PIV3. There was no clear association between infection with any of the viruses isolated or tested for and the presence of overt clinical signs of respiratory disease. CONCLUSIONS: There was serological and/or virological evidence that EHV-1, EHV-2, EHV-5, EAdV-1 and ERBV infections were present among foals in New Zealand. EHV-2 infection was first detected in foals as young as 3 months of age. The isolation of EHV-2 from nasal swabs preceded serological evidence of infection with other respiratory viruses, suggesting that EHV-2 may predispose foals to other viral infections.     

微小隐孢子虫端粒序列及端粒酶活性的研究

为了探讨我国微小隐孢子虫(C.parvum)的端粒序列,试验根据国外已报道的端粒重复序列5’-CCTAAA-3’设计寡聚核苷酸探针(CCTAAA)5,并用地高辛标记,C.parvum基因组DNA经Bal31-HindⅢ酶切后进行Southern印迹杂交鉴定;并在此基础上,采用改进的端粒重复序列扩增(TRAP)法首次对子孢子和卵囊时期C.parvum的端粒酶活性进行检测。结果表明:C.parvum基因组DNA与探针(CCTAAA)5杂交,获得了较好的杂交条带,随着Bal31-HindⅢ酶切时间的延长,杂交信号逐渐减弱,说明C.parvum端粒序列定位在染色体的末端,与国外报道的C.parvum端粒序列一致,为5’-CCTAAA-3’;检测C.parvum端粒酶活性时发现,子孢子时期端粒酶具有活性,卵囊中未检测到端粒酶活性。     

Pharmacokinetics of once-daily amikacin in healthy foals and therapeutic drug monitoring in hospitalized equine neonates

The objectives of this study were to investigate the pharmacokinetics of once-daily amikacin in healthy neonates, to determine amikacin concentrations in hospitalized foals, and to determine the minimum inhibitory concentrations (MICs) of amikacin against gram-negative isolates from blood cultures in septic foals. Median half-life, clearance, and volume of distribution of amikacin in healthy 2- to 3-day-old foals after administration of an intravenous bolus of amikacin (25 mg/kg) were 5.07 hours (4.86-5.45 hours), 1.82 mL/min/kg (1.35-1.97 mL/min/kg), and 0.785 L/kg (0.638-0.862 L/kg), respectively. Statistically significant (P <.05) decreases in area under the curve (14% decrease), mean residence time (19% decrease), and C24h plasma amikacin concentrations (29% decrease) occurred between days 2-3 and 10-11. Plasma amikacin concentrations in healthy foals at 0.5 hours (C0.5h) were significantly higher (P = .02) than those of hospitalized foals. Sepsis, prematurity, and hypoxemia did not alter amikacin concentrations. The MIC at which 90% of all gram-negative isolates from equine neonatal blood cultures were inhibited by amikacin was 4 microg/mL, suggesting that amikacin C0.5h of 40 microg/mL should be targeted to achieve a maximum serum concentration to MIC ratio of 10:1. The proportion of foals with C0.5h 40 microg/mL was significantly higher (P < .0001) in hospitalized foals receiving a dose of amikacin at 25 mg/kg (22/24 or 92%) than in foals receiving a dose at 21 mg/kg (9/25 or 36%), whereas no difference was found in the proportion of foals with C24h concentrations > or = 3 microg/mL between the 2 groups. An initial dose at 25 mg/kg is recommended for once-daily amikacin in equine neonates.     

Quantitative Risk Assessment for Zoonotic Transmission of Cryptosporidium parvum Infection Attributable to Recreational Use of Farmland

Cryptosporidiosis caused by Cryptosporidium parvum infection is a major cause of enteric illness in man and there is a significant reservoir in animals, particularly young ruminant species. To preliminary assess the magnitude of the risk posed by contact with faeces produced by infected livestock, two microbiological risk assessments have been developed: one for the risk of human infection with C. parvum while camping on contaminated land recently grazed by infected suckler cattle and a comparable risk assessment for camping on land recently spread with contaminated cattle slurry. Using a worst‐case scenario approach, the upper level of risk was estimated to be one infection in every 6211 person‐visits for a camping event on land recently grazed by infected cattle. Translated into camping events of 100 persons, this risk estimate would most likely lead to zero (98% likelihood) or one infection (1% likelihood). The results for cattle slurry model are similar despite different pathways. Sensitivity analysis was conducted for the grazing cattle model only. This suggested that the time between grazing and camping was the most important control strategy, but increasing hand‐washing frequency and the removal of cattle faeces before camping would also be beneficial. If the upper level of risk were to be judged unacceptable then further data would be required to more accurately estimate the risk of infection through these scenarios. Further research would also be required to assess the fraction of cases attributable to camping and/or environmental contact with Cryptosporidium oocysts.     

安徽省猪隐孢子虫病流行病学调查

为了查明安徽省猪隐孢子虫感染与地理分布情况,选取该省10个县、市进行了猪隐孢子虫感染情况的调研。结果在58头猪的粪样中查到了隐孢子虫卵囊,其感染率为10.05%(58/577)。经鉴定,所获虫体为小隐孢子虫(Cryptosporidium parvum)。该省猪的隐孢子虫感染存在年龄和地区性差异,但与性别无关。     

Antimicrobial therapy in neonatal foals

There are several differences in treatment of neonatal foals with antimicrobials, compared to mature horses. Firstly, the dose of many antimicrobials is different in the foal. For orally administered drugs, this may also affect their efficacy, due to different enteral absorption. Secondly, neonatal foals are not yet hindgut fermentors and this allows antimicrobials with a high propensity to cause colitis in mature horses to be used. Thirdly, toxicities are different and some antimicrobials used in mature horses, such as enrofloxacin, are not suitable for use in foals. Foal-specific information is therefore needed for their safe and effective treatment with antimicrobials.     

Musculoskeletal disorders in neonatal foals

Angular limb deformities are not uncommon in foals. Mild angular deviation due to laxity of supporting soft tissues often resolves spontaneously. However, external splinting or casting may be needed in severe cases or in those that do not resolve. When incomplete ossification of carpal or tarsal bones is the cause of the limb deformity, external support is mandatory to prevent further deformation and abnormal development of the bones. When epiphyseal and metaphyseal abnormalities cause axial deviation, surgical intervention is usually necessary. Circumferential periosteal transection and/or transphyseal bridging are methods used. The choice is dictated by the type and severity of the deformity. Flexor contractures of the forelimb vary greatly in degree and joints affected. Physical therapy combined with intermittent splint application is often successful, but surgical intervention may be necessary in unresponsive cases. Flexor tendon laxity is usually self-correcting but physical therapy, restricted exercise, and splinting may be needed. Rotational abnormalities are easier to correct in the forelimbs than in the hind limbs. Correction is usually accomplished by frequent corrective hoof trimming. Miscellaneous anomalies of the musculoskeletal system may sometimes be amenable to surgical correction, although the potential disadvantages must be carefully considered. Septic arthritis is a frequent sequela to neonatal septicemia and must be treated aggressively and early in its development. Appropriate systemic antibiotics, joint lavage, and rest are indicated. Neonatal osteomyelitis has a poor prognosis and requires prompt, vigorous therapy; even then, growth anomalies of the limb or contiguous septic arthritis may develop and further worsen the prognosis. Early accurate diagnosis and prompt appropriate therapy are vital in treating musculoskeletal disorders in foals, especially when a successful outcome is judged by the animal becoming a functional athlete.     

牛微小隐孢子虫表面抗原CP15基因原核表达及其表达产物的鉴定

试验旨在构建微小隐孢子虫表面抗原CP15基因原核表达载体并获得重组CP15蛋白。试验以已知重组质粒pMD-CP15为模板,PCR方法扩增出CP15基因片段,并亚克隆到pGEX4T-3,构建了在E.coli BL21中GST融合表达载体pGEX-CP15;经1 mmol/L IPTG进行诱导表达获得目的蛋白,其大小采用SDS-PAGE电泳和Western blotting进行鉴定。结果表明,扩增出约390 bp的微小隐孢子虫表面抗原CP15基因片段并成功构建pGEX-CP15质粒,表达出分子质量为42 ku的融合蛋白,与推导的理论值相符。Western blotting显示该蛋白能被GST血清识别。牛微小隐孢子虫表面抗原CP15基因在大肠杆菌中得到了高效表达。     

The occurrence of Cryptosporidium parvum,Campylobacter and Salmonella in newborn dairy calves in the Manawatu region of New Zealand

AIM: To determine the occurrence of Cryptosporidium parvum oocysts, Campylobacter spp and Salmonella spp in faecal samples taken from newborn dairy calves on 24 dairy farms in the Manawatu region of New Zealand.

METHODS: A cross-sectional study was conducted during the 2002 calving season. Faecal samples were collected from 185 newborn calves from a convenience sample of 24 dairy farms. The samples were tested microscopically for the presence of C. parvum oocysts, and bacteriologically for the presence of Campylobacter spp and Salmonella spp.

RESULTS: Infections with C. parvum were identified in 33/156 (21.2%) calves from 10 farms. More than 10⁶ oocysts/g (OPG) faeces were detected in calves from four farms. Campylobacter spp were isolated from 58/161 (36%) calves from 18 farms; in particular, C. jejuni subsp jejuni was isolated from 11/161 (6.8%) calves from seven farms. Salmonellae were not detected.

CONCLUSIONS: Despite the short and concentrated calving pattern and the long interval between calving seasons characterising most dairy farms in New Zealand, C. parvum is widespread among calves. Campylobacter spp, especially C. jejuni, rapidly colonise the intestinal tract of newborn calves.

RELEVANCE: This study provided an estimate of the ecological impact of newborn dairy calves with regard to the potentially zoonotic enteric pathogens most frequently isolated from human gastrointestinal infections in New Zealand.     

Comparison of blood electrolyte and biochemical parameters between single infections of rotavirus and Cryptosporidium parvum in diarrheic Hanwoo calves

BackgroundNeonatal calf diarrhea is a major problem in the cattle industry worldwide. Rotavirus and Cryptosporidium parvum are the primary causative agents, especially during the first three weeks of the calf’s life.ObjectivesThis study investigated the differences in acid-base, electrolytes, and biochemical parameters of diarrheic calves with infection of either rotavirus or C. parvum.MethodsA total of 61 Korean native calves (≤ 20 days old) were divided into two groups based on rotavirus or C. parvum infections: rotavirus infection (n = 44) and C. parvum infection (n = 17). The calves with at a specific blood pH range (pH 6.92–7.25) were chosen for comparison. The acid-base, electrolyte, chemistry, and serum proteins were analyzed, Further, fecal examinations were performed.ResultsCompared to C. parvum-infected calves, the rotavirus-infected calves showed lower levels of total carbon dioxide, bicarbonate (HCO₃ ⁻), anion gap, total protein, and albumin/globulin ratio, and significantly lower levels of potassium, globulin, and α2-globulin (p < 0.05). The C. parvum-infected calves (r = 0.749) had stronger correlations between pH and HCO₃ ⁻ than the rotavirus-infected calves (r = 0.598). Compared to rotavirus-infected calves, strong correlations between globulin and α2-globulin, α2-globulin and haptoglobin were identified in C. parvum-infected calves.ConclusionsThis study is the first to investigate acid-base, electrolyte, and biochemical parameters in calves in response to infections of rotavirus and C. parvum. Although rotavirus and C. parvum cause malabsorptive and secretory diarrhea in similar-aged calves, blood parameters were different. This would help establish the diagnostic and treatment strategies.     

以CP23重组蛋白为抗原建立检测微小隐孢子虫抗体间接ELISA方法的研究

摘要：以在E.coli高效表达的微小隐孢子虫子孢子表面抗原CP23为抗原，以辣根过氧化物酶(HRP)标记的山羊抗鼠IgG为二抗，建立了检测微小隐孢子虫抗体的间接ELISA方法。经检测筛选出最佳反应条件为1μg／孔纯化的E.coli表达的CP23抗原包被酶标板，用10％免血清进行封闭，以正常E.coli裂解上清液稀释待检血清。实验表明应用CP23重组蛋白作为诊断C.parvum抗原具有特异性高、抗原易纯化和成本低等特点。     

微小隐孢子虫抗原CTL细胞表位预测及多表位基因的原核表达

应用多个服务器对微小隐孢子虫（Cryptosporidium parvum）候选疫苗抗原的氨基酸序列进行分析,预测可能存在的CD8＋细胞毒性T细胞（CD8＋cytotoxic T lymphocyte,CD8＋CTL）表位。从6种常见的免疫效果较好的候选疫苗抗原基因中选出10个分值较高的表位基因串联在一起,形成一条多表位基因,进行人工合成,表位之间用柔性氨基酸GGGGS或GPGPG链接,命名为CpCTL10。将该多表位基因重组入高效融合表达载体pET-28a（＋）,获得重组质粒pET-28a（＋）-CpCTL10,转化大肠杆菌BL21（DE3）进行诱导表达,表达产物进行SDS—PAGE、Western blot分析。结果显示,CpCTL10基因在大肠杆菌中主要以包涵体形式高效表达,表达的重组蛋白占菌体蛋白总量的55．3％,纯化的重组蛋白纯度达75．1％。Western blot分析显示表达产物能与感染隐孢子虫的鼠阳性血清发生特异性反应,表明表达的重组蛋白具有较好的抗原性,为多抗原多表位疫苗的研制打下某础．     

小球隐孢子虫(Cryptosporidium parvum)P15基因的克隆和表达

通过PCR扩增出小球隐孢子虫(Cryptosporidium parvum)P15基因片段。将该片段连接到克隆质粒裁体pUC19的BamH Ⅰ酶切位点上，并对其序列进行了测定。结果表明，该基因长度为479bp，编码的表面蛋白由148个氨基酸残基组成。再将P15蛋白基因片段分别在大肠杆菌和哺乳动物细胞中表达，经SDS-PAGE分析和Western blot检测，结果表明，小球隐孢子虫P15蛋白基因在大肠杆菌内获得表达的融合蛋白相对分子质量为50000；在哺乳动物细胞中，重组痘病毒表达的蛋白相对分子质量为18000-22000，与直接从自然感染牛小球隐孢子虫分离的P15蛋白的相对分子质量相似。