Role of Campylobacter jejuni potential virulence genes in cecal colonization

Campylobacter jejuni, a common commensal in chickens, is one of the leading causes of bacterial gastroenteritis in humans worldwide. The aims of this investigation were twofold. First, we sought to determine whether mutations in the C. jejuni ciaB and pldA virulence-associated genes impaired the organism's ability to colonize chickens. Second, we sought to determine if inoculation of chicks with C. jejuni mutants could confer protection from subsequent challenge with the C. jejuni wild-type strain. The C. jejuni ciaB gene encodes a secreted protein necessary for the maximal invasion of C. jejuni into cultured epithelial cells, and the pldA gene encodes a protein with phospholipase activity. Also included in this study were two additional C. jejuni mutants, one harboring a mutation in cadF and the other in dnaJ, with which we have previously performed colonization studies. In contrast to results with the parental C. jejuni strain, viable organisms were not recovered from any of the chicks inoculated with the C. jejuni mutants. To determine if chicks inoculated with the C. jejuni mutants become resistant to colonization by the C. jejuni parental strain upon subsequent challenge, chicks were inoculated either intraperitoneally (i.p.) or both orally and i.p. with the C. jejuni mutants. Inoculated birds were then orally challenged with the parental strain. Inoculation with the C. jejuni mutants did not provide protection from subsequent challenge with the wild-type strain. In addition, neither the C. jejuni parental nor the mutant strains caused any apparent morbidity or mortality of the chicks. We conclude that mutations in genes cadF, dnaJ, pldA, and ciaB impair the ability of C. jejuni to colonize the cecum, that chicks tolerate massive inoculation with these mutant strains, and that such inoculations do not provide biologically significant protection against colonization by the parental strain.     

Food of Canis Mesomelas in South Africa

Data from five jackass penguin chicks (mean mass 201 g) showed that even young chicks can increase their oxygen consumption from 0,76± 0,16 ml 0¹/gxhr at 35°C to 2,09± 0,21 ml 0, at 20 °C. Close parental attendance may therefore be interpreted not only as protection against aerial predators but also as creating a microclimate which could favour a decrease in maintenance requirements and thus channel a substantial part of the available energy into growth. A decrease in rectal temperature, despite the increase in heat production, is interpreted as indicative of poor insulative properties of the down. Preliminary data on the ontogeny of thermoregulation indicate that penguin chicks may attain mature thermoregulatory abilities at a mass of approximately 400g.     

Acclimatization of the fowl to intermittent acute heat stress

The acclimatization of poultry to a hot, dry climate was assessed by studying changes in rectal temperature (Tr) during regular daily exposure to an ambient temperature of 38 degrees C and 26% RH. Laying hens of two strains, young chicks, growing broilers and broiler breeders were able to acclimatise, in varying degrees, to these conditions. Acclimatization was characterised by a progressive reduction in the rate of increase in Tr over the period of exposure and the ability to survive conditions that initially would have been fatal. Laying hens could acclimatise if prevented from displaying their normal postural responses to heat stress. Acclimatization was accompanied by a decrease in oxygen consumption when measured at either the acclimatization temperature or at a thermoneutral temperature.     

ORIGINAL ARTICLE: Ileal endogenous amino acid flow of broiler chickens under high ambient temperature

High environmental temperature has detrimental effects on the gastrointestinal tract of poultry. An experiment was conducted to determine the effect of acute heat stress on endogenous amino acid (EAA) flow in broiler chickens. A total of 90, day‐old broiler chicks were housed in battery cages in an environmentally controlled chamber. Chicks were fed a nitrogen‐free diet on day 42 following either no heat exposure (no‐heat) or 2 weeks exposure to 35 ± 1 °C for 3 h from days 28 to 42 (2‐week heat) or 1 week exposure to 35 ± 1 °C for 3 h from days 35 to 42 (1 week heat). The most abundant amino acid in the ileal flow was glutamic acid, followed by aspartic acid, serine and threonine in non‐heat stressed group. The EAA flow in 1‐week heat and 2‐week heat birds were significantly (p < 0.05) higher than those under no heat exposure (14682, 11161 and 9597 mg/kg of dry matter intake respectively). Moreover, the EAA flow of 2‐week heat group was less than 1‐week heat group by approximately 36%. These observations suggest that the effect of heat stress on EAA flow is mostly quantitative; however, heat stress may also alter the content of EAA flow qualitatively.     

Effects of electroanesthesia and a phenothiazine tranquilizer on thermoregulation in the sheep.

The effects of giving propiopromazine alone and of electroanesthesia-propiopromazine treatment on thermoregulation (body temperature regulation) were studied in 3 sheep at ambient temperatures of 5, 25, and 35 C. Measures of thermoregulation during a 120-minute treatment and 120-treatment recovery period included rectal temperature, respiratory frequency, respiratory evaporative heat loss, metabolic heat production, multiple skin temperatures, and shivering. During cold exposure (5 C), both the propiopromazine administration and the electroanesthesia-propiopromazine treatment resulted in hypothermia which was attributed to increased peripheral and respiratory heat losses, a transient inhibition of shivering thermogenesis, and a reduction in metabolic heat production. At 35 C ambient temperature, both resulted in hyperthermia caused principally by a reduction in respiratory evaporative heat loss. The effects of electroanesthesia-propiopromazine treatment on thermoregulation appeared to be additive at both the cold (5 C) and the hot (35 C) environments, in that simultaneous administration resulted in a more profound thermoregulatory impairment. Nevertheless, shifts in body temperature during electroanesthesia are partly attributable to phenothiazine premedication.     

Chicken anemia agent in the United States: isolation of the virus and detection of antibody in broiler breeder flocks

Chicken anemia agent (CAA) was isolated from broiler chickens in Texas with a blue wing or anemia dermatitis-like syndrome. Specific-pathogen-free chicks inoculated with field material developed anemia, and CAA was isolated in MDCC-MSB1 cells from bone marrow and lymphoid tissue from inoculated chicks. One isolate, designated EF88/78/276, was further characterized. Infectivity of EF88/78/276 was resistant to treatment with chloroform and with heat at 70 C for 5 minutes. EF88/78/276 was indistinguishable from the Cux-1 and Gifu-1 isolates of CAA by cross-neutralization tests. Almost all 1-day-old susceptible chicks inoculated intramuscularly with EF88/78/276 developed anemia, but contact-infected chicks did not. Antibody to CAA was detected in broiler breeder flocks from Texas, the Delmarva peninsula, and Alabama.     

Comparison of heat production and plasma lipid metabolites between meat- and egg-types of Nagoya breed chicken during embryonic development

We compared heat production (HP) and plasma lipid metabolites between meat‐ and egg‐types of Nagoya breed chickens during embryonic development. To investigate HP and respiratory quotient, oxygen consumption and carbon dioxide production were measured using an open‐circuit calorimeter system. HP was significantly lower in meat‐ than in egg‐type chickens during embryonic development, and HP gradually decreased with developmental stage in both types. The respiratory quotient was constant at approximately 0.68 at every embryonic stage investigated, and the value was similar in both types. Plasma concentrations of triacylglycerol, nonesterified fatty acid, glycerol and D‐3‐hydroxybutyrate, which are associated with lipid metabolism, were similar in both types during embryonic development. These results demonstrate that chicken embryos selected for rapid growth of Nagoya breed have characteristic lower HP, and that when selecting chickens for rapid growth, HP is an important parameter during embryonic stages.     

持续高温对不同性别北京油鸡脂肪代谢及血液脂肪细胞因子水平的影响

本研究旨在考察持续高温对不同性别北京油鸡脂肪代谢及血液脂肪细胞因子水平的影响.选择健康、体重相近的49日龄北京油鸡公母各72只[公鸡平均体重(608.58±13.07)g;母鸡平均体重(518.50±8.33)g],随机分在2个处理中:高温组,(35.00±0.37)℃;适温组,(25.00±0.38)℃,每个处理设6个重复,每个重复6只鸡.试验持续23 d.测定试验第12、23天时的血液生化指标、血液脂肪细胞因子水平及脂肪组织脂蛋白酯酶活性.结果表明:1)对公鸡而言:持续高温12 d时血清胰岛素水平显著升高(P＜0.01),血浆甘油三酯含量显著降低(P＜0.01).持续高温23 d时血清胰岛素水平和血浆甘油三酯含量显著降低(P＜0.01),血浆葡萄糖含量显著升高(P＜0.01);对母鸡而言:持续高温12 d时血浆甘油三酯、游离脂肪酸含量显著降低(P＜0.05).持续高温23 d时血浆甘油三酯、游离脂肪酸含量无显著变化(P＞0.05),血浆极低密度脂蛋白含量有增加趋势(P =0.065),血浆葡萄糖含量显著升高(P＜0.01).2)持续高温12 d时公母鸡的肌间脂肪脂蛋白酯酶活性都显著降低(P＜0.05),持续高温23 d时公鸡的肌间脂肪脂蛋白酯酶活性显著降低(P＜0.05),而母鸡的无显著变化(P＞0.05).3)公母鸡血浆脂联素、抵抗素水平在高温下都无显著变化(P＞0.05);持续高温12 d时公鸡血浆瘦素水平显著降低(P＜0.01),母鸡的显著升高(P＜0.05).由此可知,随着持续高温时间的延长,母鸡血浆游离脂肪酸、甘油三酯含量及肌间脂肪组织脂蛋白酯酶活性受高温影响下降程度减小,公鸡血浆甘油三酯含量及脂肪组织脂蛋白酯酶活性下降程度加大.     

Single nucleotide polymorphisms associated with thermoregulation in lactating dairy cows exposed to heat stress

Dairy cows with increased rectal temperature experience lower milk yield and fertility. Rectal temperature during heat stress is heritable, so genetic selection for body temperature regulation could reduce effects of heat stress on production. One aim of the study was to validate the relationship between genotype and heat tolerance for single nucleotide polymorphisms (SNPs) previously associated with resistance to heat stress. A second aim was to identify new SNPs associated with heat stress resistance. Thermotolerance was assessed in lactating Holsteins during the summer by measuring rectal temperature (a direct measurement of body temperature regulation; n = 435), respiration rate (an indirect measurement of body temperature regulation, n = 450) and sweating rate (the major evaporative cooling mechanism in cattle, n = 455). The association between genotype and thermotolerance was evaluated for 19 SNPs previously associated with rectal temperature from a genomewide analysis study (GWAS), four SNPs previously associated with change in milk yield during heat stress from GWAS, 2 candidate gene SNPs previously associated with rectal temperature and respiration rate during heat stress (ATPA1A and HSP70A) and 66 SNPs in genes previously shown to be associated with reproduction, production or health traits in Holsteins. For SNPs previously associated with heat tolerance, regions of BTA4, BTA6 and BTA24 were associated with rectal temperature; regions of BTA6 and BTA24 were associated with respiration rate; and regions of BTA5, BTA26 and BTA29 were associated with sweating rate. New SNPs were identified for rectal temperature (n = 12), respiration rate (n = 8) and sweating rate (n = 3) from among those previously associated with production, reproduction or health traits. The SNP that explained the most variation were PGR and ASL for rectal temperature, ACAT2 and HSD17B7 for respiration rate, and ARL6IP1 and SERPINE2 for sweating rate. ARL6IP1 was associated with all three thermotolerance traits. In conclusion, specific genetic markers responsible for genetic variation in thermoregulation during heat stress in Holsteins were identified. These markers may prove useful in genetic selection for heat tolerance in Holstein cattle.     

Cell-mediated and humoral immune responses to a virulent plasmid-cured mutant strain of Salmonella enterica serotype gallinarum in broiler chickens

One-week-old Salmonella-free broiler chicks were subcutaneously immunized and subsequently boosted two weeks later with 2x10(7)cfu 0.5ml(-1) of an 85-kb virulent plasmid-cured spectinomycin-resistant mutant strain (SG9VP(-)Spc(r)) of Salmonella gallinarum 9 (S. enterica serotype gallinarum 9) along with a control group of mock-immunized chickens. The chicks were subcutaneously challenged at 5 weeks of age with 5x10(10)cfu 0.5ml(-1) (5xLD(50)) of wild-type S. gallinarum var. Duisburg (S. enterica serotype gallinarum var. Duisburg). The cellular and humoral immune responses were measured at weekly intervals post-immunization (PI) and post-challenge (PC) using lymphocyte stimulation test (LST), delayed type hypersensitivity (DTH) test, serum tube agglutination test (STAT) and enzyme-linked immunosorbent assay (ELISA). High stimulation indices suggestive of a potent lymphocyte transformation response and high persistent serum IgG titres were recorded in immunized chickens at the termination of the experiment. These findings indicate that the live attenuated mutant vaccine induced a strong cellular and humoral immunity, which may play a role in the protection of fowl typhoid in broiler chickens.     

热应激状态下羊的体温调节

热应激是由高温环境所引起的,在炎热的气候条件下,当环境有效温度超过羊的等热区上限温度时,羊就会出现热应激。由热应激引发的不良影响广泛存在,不仅严重影响羊的生存质量,而且导致羊体温升高、生长性能降低、免疫力减弱以及肉品质下降。本文综述了导致羊产生热应激的因素以及热应激条件下的体温调节机制。在热应激的情况下,机体通过吸收环境辐射热和累积机体代谢产热,使机体获得很高的热负荷,超过了机体的散热能力。暴露在高温环境中的羊只,可通过增加机体散热消除过多的热负荷。当空气温湿度发生变化时,作为恒温动物,绵羊和山羊可以通过调整自身的产热量、采食量、饮水量、呼吸频率和行为等方式,来维持机体产热和散热的平衡。总之,羊可以利用复杂有效的散热机制维持体温平衡,适应高温环境。     

早期热应激对肉仔鸡后期生长发育的影响

旨在研究早期热应激对肉仔鸡后期生长发育及骨骼肌中相关信号通路基因表达的影响。本研究选取健康、体重相近的刚出壳AA肉仔鸡192只（公母各半）,随机分成2组（每组4个重复,每个重复24只鸡）,饲喂至3日龄时,分别给予36℃热应激处理24 h （热应激组）和正常温度（33℃）饲喂（对照组）。42日龄时每个处理选取8只鸡采取血浆,屠宰后测定胴体性能,分离胸肌样品测定骨骼肌发育有关的基因表达。结果表明,热应激当天肉仔鸡增重显著降低,但整个饲养期肉仔鸡的平均日增重显著增加（P<0.05）;与对照组相比,早期热应激有提高整个饲养期肉仔鸡平均日采食量的趋势（0<P<0.1）。早期热应激对肉仔鸡的屠宰率、半净膛重、全净膛重、肝比重、心比重、胸肌率、腿肌率、腹脂率都无显著影响（P>0.05）。早期热应激显著升高了42日龄肉仔鸡血浆酮体含量（P<0.05）。在胸肌中,早期热应激显著提高了胰岛素样生长因子（IGF-1）及其受体（IGF-1R）的基因表达（P<0.05）。本试验结果提示,早期热应激促进了肉仔鸡后期的生长发育,骨骼肌中IGF-1信号可能参与此调节过程。     

Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat‐stressed broiler chickens and its effect on heat shock protein 70 expression,blood parameters and body temperature

The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat‐stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35–42, heat stress was induced by exposing birds to 38 ± 1 °C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress.     

The effects of early‐age thermal manipulation and daily short‐term fasting on performance and body temperatures in broiler exposed to heat stress

This study was conducted to investigate the effects of thermal manipulation at 5 days of age and short‐term fasting during the warmest part of the day on responses to prolonged heat stress of broilers. A total of 240‐day‐old Ross 308 female broiler chicks were divided into three groups: control, thermal manipulation (chicks were exposed to 36 °C for 24 h at 5 days of age) and short‐term fasting during the warmest part of the day (10.00–17.00 h). Prolonged heat stress was induced daily from 28 to 42 days by heating until the ambient temperature reached 32–35 °C between 10.00 and 17.00 h. Both thermal manipulation and short‐term fasting resulted in a decrease in rectal temperatures and haematocrit values at 35 and 41 days of age. Thermal manipulation improved body weight, feed consumption and feed conversion. However, short‐term fasting caused a reduction in body weight and a deterioration in feed conversion. Short‐term fasting lowered the percentages of carcass, whereas thermal manipulation highered breast yield. Both thermal manipulation and short‐term fasting decreased heart mass and abdominal fat.     

The absence of cecal colonization of chicks by a mutant of Campylobacter jejuni not expressing bacterial fibronectin-binding protein.

Campylobacter jejuni is a common cause of human gastrointestinal illness throughout the world. Infections with C. jejuni and Campylobacter coli are frequently acquired by eating undercooked chicken. The ability of C. jejuni to become established in the gastrointestinal tract of chickens is believed to involve binding of the bacterium to the gastrointestinal surface. A 37-kD outer membrane protein, termed CadF, has been described that facilitates the binding of Campylobacter to fibronectin. This study was conducted to determine whether the CadF protein is required for C. jejuni to colonize the cecum of newly hatched chicks. Day-of-hatch chicks were orally challenged with C. jejuni F38011, a human clinical isolate, or challenged with a mutant in which the cadF gene was disrupted via homologous recombination with a suicide vector. This method of mutagenesis targets a predetermined DNA sequence and does not produce random mutations in unrelated genes. The parental C. jejuni F38011 readily colonized the cecum of newly hatched chicks. In contrast, the cadF mutant was not recovered from any of 60 chicks challenged, indicating that disruption of the cadF gene renders C. jejuni incapable of colonizing the cecum. CadF protein appears to be required for the colonization of newly hatched leghorn chickens.     

Characterization of a novel nicotinamide adenine dinucleotide-cytochrome b5 reductase mutation associated with canine hereditary methemoglobinemia

Hereditary methemoglobinemia associated with nicotinamide adenine dinucleotide-cytochrome b5 reductase (b5R) deficiency is a rare autosomal recessive disorder in animals. Recently, nonsynonymous b5R gene (CYB5R3) variants have been reported to be associated with canine and feline hereditary methemoglobinemia. However, the underlying molecular mechanisms of canine and feline methemoglobinemia caused by these nonsynonymous variants have not yet been reported. Previously, we reported a Pomeranian dog family with hereditary methemoglobinemia, carrying CYB5R3 mutation of an A>C transition at codon 194 in exon 7, replacing an isoleucine residue with leucine (p.Ile194Leu). In this study, we investigated the enzymatic and structural properties of the soluble form of wild-type and Ile194Leu canine b5Rs to characterize the effects of this missense mutation. Our results showed that the kinetic properties of the mutant enzyme were not affected by this amino acid substitution. The secondary structure of the wild-type and Ile194Leu b5Rs detected by circular dichroism showed a similar pattern. However, the mutant enzyme exhibited decreased heat stability and increased susceptibility to trypsin hydrolysis. Moreover, the thermostability and unfolding measurements indicated that the mutant enzyme was more sensitive to temperature-dependent denaturation than the wild-type b5R. We concluded from these results that unstable mutant enzyme properties with normal enzymatic activity would be associated with hereditary methemoglobinemia in the Pomeranian dog family.     

Effect of metC mutation on Salmonella Gallinarum virulence and invasiveness in 1-day-old White Leghorn chickens

Salmonella enterica serotype Gallinarum (S. Gallinarum) is the causative agent of fowl typhoid (FT) in chickens. FT is a severe systemic disease of chickens causing heavy economic losses to the poultry industry through mortality, reduced egg production and culling of precious breeding stocks. In this study, a metC (encoding cystathionine beta lyase) mutant was produced from a virulent strain of S. Gallinarum by Mini-Tn5 insertional inactivation. The mutant was significantly attenuated in virulence for 1-day-old White Leghorn chickens. Inactivation of metC resulted in 10(4)-fold increase in the LD50 when compared with the wild type parent. The metC mutant showed an in vivo competitiveness defect in the challenged chickens and significantly lower (P < 0.01) bacterial burden in the reticuloendothelial organs when compared with the wild-type parent. These results indicate that metC gene is important for virulence of S. Gallinarum in chickens.     

益生菌和葡聚糖对冷应激状态下贵妃雏鸡血液学变化的影响

选择1日龄健康贵妃雏鸡192只,随机分为16组,每组12只。试验雏鸡经受比正常温度低10℃的急、慢性冷应激和冷适应处理,并对急性冷应激组添加益生菌、慢性冷应激和冷适应组添加葡聚糖处理。翼下采血制血涂片,镜检观察雏鸡白细胞分类变化,并用血细胞分析仪测定血相变化与之对比分析。结果表明,在急性冷应激期间,雏鸡的白细胞分类计数波动变化明显,嗜碱粒细胞数目和H/L比值都明显增加;血红蛋白、红细胞以及血小板数目显著增加。冷适应后淋巴细胞数目明显增加,嗜中性粒细胞明显减少,H/L比值下降。益生菌、葡聚糖处理对白细胞总数和嗜酸粒细胞影响明显,益生菌处理24h明显增加白细胞数量,葡聚糖处理15d降低了嗜酸粒细胞数目。本试验结果提示急性冷应激能引起雏鸡血液学参数的明显波动,贵妃雏鸡抗冷应激能力较强,而益生菌、葡聚糖能提高雏鸡适应冷应激的能力,改善应激状态下的免疫功能。     

Relative humidity at moderate ambient temperatures: its effect on male broiler chickens and turkeys

1. The effect of relative humidity (RH, 40% to 75%) at moderate ambient temperatures (Ta, 28 degrees and 30 degrees C) on the performance and thermoregulation of male broiler chickens and turkeys was studied at the age of 4 to 8 weeks. 2. Weight gain and food intake of male broiler chickens were significantly higher at 60% to 65% RH but food conversion efficiency was not affected by RH. In male turkeys, no effect of RH on performance was recorded. 3. Both chickens and turkeys controlled body temperature at normothermic levels during exposure to the experimental environmental conditions. 4. The rate of panting estimated from blood pH and pCO2 was lower in chickens exposed to 28 degrees C than to 30 degrees C. In turkeys, the rate was lower than that recorded in chickens at both Ta(s). 5. Plasma T3 was positively and significantly correlated with food intake. 6. It may be concluded that RH plays a role in the performance of chickens exposed to 30 degrees and 28 degrees C. whereas male turkeys must respond to RH at Ta >30 degrees C.     

The effect of early-age food restriction on heat shock protein 70 response in heat-stressed female broiler chickens

1. This study was conducted to determine the effect of early-age food restriction on heat shock protein (hsp) 70 synthesis in the brains of female broiler chickens exposed to high ambient temperatures. 2. Chicks were brooded for 3 weeks and then maintained at 24+/-1 degrees C. 3. On d 0, chicks were assigned to one of 4 feeding regimens; each regimen was applied to 4 cages of chicks. The regimens were: (1) ad libitum feeding (AL); (2) 80% food restriction at 4, 5 and 6 d of age (F80); (3) 60% food restriction at 4, 5, and 6 d of age (F60); and (4) 40% food restriction at 4, 5 and 6 d of age (F40). From d 35 to d 41, all chicks were subjected to 38+/-1 degrees C for 2 h/d. 4. One day following food restriction (d 7), hsp 70 expression in the brain samples of F60 and F40 chicks was augmented but not those fed AL and F80. 5. Prior to the heat challenge (d 35), all chicks had similar hsp 70 response. Irrespective of feeding regimen, there was a marked increase in hsp 70 expression after 4 d of heat treatment (d 38). Following 7 d of heat exposure (d 41), except for the F60 chicks, the augmented hsp 70 expression in the brains of AL, F80 and F40 birds was not maintained. 6. Enhancement of hsp 70 expression was noted in birds subjected to F60, but not AL, F80 or F40, throughout the period of heat exposure.