In vivo screening of four phytochemicals/extracts and a fungal immunomodulatory protein against an Eimeria acervulina infection in broilers

Background: Besides the anticoccidial drug resistance problem, increasing consumer concerns about food safety and residues have propelled the quest for alternative prevention and control strategies amongst which phytotherapy has gained appeal due to a renewed interest in natural medicine.Objective: The objective was in vivo screening of four phytochemicals/extracts and a fungal immunomodulatory protein (FIP) against an Eimeria acervulina infection in broilers.Animals and methods: Four phytochemicals/extracts (extract from Echinacea purpurea, betaine (Betain?), curcumin, carvacrol (two different doses)), and a recombinant FIP from Ganoderma lucidum cloned and expressed in Escherichia coli were investigated for their anticoccidial potential. The experiment was conducted in a battery cage trial with 54 cages of eight birds each. Broilers infected with E. acervulina (a low and high infection dose of 10⁴ and 10⁵ sporulated oocysts, respectively) and treated with the phytochemicals/extracts or the FIP were compared with broilers treated with the anticoccidial salinomycin sodium (Sacox®) and with an untreated uninfected and an untreated infected control group. Coccidiosis lesion scores, body weight gains and oocyst shedding were used as parameters.Results: The results showed a coccidiosis infection dose effect on the mean coccidiosis lesion scores. The phytochemicals/extracts and the FIP failed to reduce coccidiosis lesion scores and oocyst shedding, while salinomycin efficiently controlled the E. acervulina infection and enabled significantly higher body weight gains.Conclusion: In conclusion, the selected phytochemicals/extracts and the FIP did not reduce the lesions of an experimentally induced E. acervulina infection.     

Three Enzymes Newly Identified from the Genus Eimeria and Two More Newly Identified from E. maxima,Leading to the Discovery of Some Aliphatic Acids with Activity against Coccidia of the Domesticated Fowl

Nine enzymes were detected in sporulated oocysts of Eimeria tenella and E. maxima, parasites of the domesticated fowl (Gallus gallus). Three enzymes, hydroxybutyrate dehydrogenase, alanine aminotransferase and -glutamyltransferase, all identified for the first time in Eimeria of fowl, occurred both in E. tenella and in E. maxima. The remaining enzymes assayed had previously been found in various Eimeria species of fowl, although creatine kinase and glutamate dehydrogenase were hitherto unknown from E. maxima. The three enzymes newly recorded from Eimeria of fowl are of interest as potential genetic markers, and also as potential chemotherapeutic targets. The discovery of hydroxybutyrate dehydrogenase led to the demonstration of anticoccidial activity by some aliphatic acids. The paper also includes a list of the enzymes detected in Eimeria of fowl in previous studies.     

Anticoccidial Vaccination of Broiler Chickens in Various Management Programmes: Relationship between Oocyst Accumulation in Litter and the Development of Protective Immunity

Paracox anticoccidial vaccine was administered to a 7-day-old flock of commercial broiler breeder stock subsequently reared to point-of-lay in the same house. For comparison, three subgroups of another flock of broiler breeders were also vaccinated with Paracox at 7 days of age, reared to 42 days and then transferred to new litter on another farm until point-of-lay. The first subgroup received no further treatment, but the second and third each received a second vaccination with Paracox, either immediately after transfer to the new litter or 42 days after transfer. Using an Eimeria necatrix model, protective immunity was demonstrated by virulent challenge of samples of birds from all groups by the age of 37–40 days (30–33 days after the first vaccination), and was maintained to at least 122–125 days of age, whether the birds remained on the same litter or were transferred to another farm, and whether they received one or two anticoccidial vaccinations. Therefore, there is no disadvantage in transferring birds onto new litter 35 days after a single Paracox vaccination, nor is there any advantage in giving a second vaccination after such a transfer. Vaccinated birds seeded the new litter with oocysts, despite being clinically immune to coccidiosis. A supplementary laboratory experiment showed that birds vaccinated at 8 days of age passed almost no oocysts after a second vaccination at 43 days of age. This indicated that they were not only protected against clinical coccidiosis, but were almost solidly immune to a homologous infection 5 weeks after a single vaccination. Nevertheless, oocysts appeared in the litter of all four groups of commercial breeders throughout the trial, showing that wild-type heterologous infections occurred whether the birds were transferred to new litter or not, but these did not overwhelm the acquired protective immunity and cause clinical coccidiosis.     

Detection of equine Babesia spp. gene fragments in Dermacentor nuttalli Olenev 1929 infesting mongolian horses,and their amplification in egg and larval progenies

Babesia equi (EMA-1) and Babesia caballi (BC48) gene fragments were amplified by polymerase chain reaction (PCR), in blood samples, and partially fed-females and egg and larval progenies of Dermacentor nuttalli, collected from horses in Altanbulag, Tuv Province, Mongolia. While Babesia parasite DNA was detected in some horse blood samples during the first PCR, all positive cases in partially fed-female ticks, eggs and larvae were confirmed by nested PCR. Present study reinforces earlier similar findings in unfed D. nuttalli ticks collected from an open space vegetation in Bayanonjuul, Tuv Province in Central Mongolia, pointing to the most likely important role of D. nuttalli in the transmission of equine babesiosis in Mongolia. The detection of parasite DNA in eggs and larval progenies is likewise suggestive of transovarial parasite transmission in this tick species.     

柔嫩艾美耳球虫感染鸡盲肠和脾脏一氧化氮合酶的表达

采用NADPH—d(nicotinamide adenine dinucleotide phosphate—diaphroase)组织化学法观察了雏鸡感染柔嫩艾美耳珠虫(E．tenella)后一氧化氮合酶(N0S)在盲肠和脾脏中的分布与表达情况。试验结果表明，所有正常鸡盲肠粘膜下层和肌层均有较深的着色，根据以往的资料和N0S的表达特性初步判断为神经元型N0S(nNOS)；试验鸡在感染后3～5d，盲肠粘膜上皮和肠腺上皮也有较深的着色，并从感染后7d开始着色减弱；而对照组鸡盲肠粘膜上皮和肠腺上皮以及对照组和试验组的脾脏几乎不着色或着色很浅。试验结果提示，盲肠粘膜上皮和肠腺上皮的着色可能是诱导型N0S(iN0S)表达的结果，而由其产生的N0参与雏鸡球虫感染过程。     

Comparative analysis of evaluation parameters in E. acervulina,E. maxima and E. tenella-infected broilers

Three parameters, body weight gain (BWG), intestinal lesion score (LS) and fecal oocyst shedding, were compared in broilers infected with major parasitic species; Eimeria acervulina, E. maxima, and E. tenella. First, two- and three-week-old chickens with Eimeria infection showed LS of approximately 3, but two-week-old chickens were more correlated with BWG. Second, significant differences in BWG were observed between male and female broilers challenged with Eimeria. Finally, E. maxima-infected broilers among three Eimeria species showed a higher relationship between BWG and LS, suggesting three considerations such as genders, age and Eimeria species for Eimeria experiments.     

Seroprevalence of Babesia bovis and Babesia bigemina in cattle in the Soutpansberg region, Limpopo Province, South Africa, associated with changes in vector-tick populations

A survey was conducted at 30 communal dip tanks and on 5 commercial farms in Limpopo Province, South Africa, during 1999 and 2000 to determine the seroprevalence of antibodies to Babesia bovis and Babesia bigemina. Cattle seropositive for B. bovis were found in 97% of the herds on communal land; the overall seroprevalence changed little between 1999 (63.3%) and 2000 (62.4%). All herds surveyed were infected with B. bigemina, and overall seroprevalence decreased significantly from 56.1% in 1999 to 49.3% in 2000. In herds on communal land in Sour Lowveld Bushveld, overall seroprevalence of B. bovis increased from 70% in 1999 to 80% in 2000, while seroprevalence of B. bigemina decreased from 70% in 1999 to 30% in 2000. This was possibly due to an influx of Rhipicephalus (Boophilus) microplus that occurred at the time. In commercially farmed herds the seroprevalence to B. bovis increased significantly from 19% in 1999 to 57.5% in 2000. All commercial herds in the survey tested positive to B. bigemina, with a seroprevalence of 48.3% in 1999 and 47.5% in 2000. During 1999, cattle in 60% of the dip tank/farm herds with only R. (B.) microplus present were approaching endemic stability to both B. bovis and B. bigemina. In 2000, 60% of the herds with only R. (B.) microplus present were approaching endemic stability for B. bovis, while only 45% were approaching endemic stability for B. bigemina. Those dip tanks/farms where only R. (B.) microplus was recorded had a significantly higher seroprevalence of B. bovis than those where both tick species were present.     

柔嫩艾美耳球虫感染雏鸡LPO含量和抗自由基酶活性的动力学

通过测定柔嫩艾美耳球虫 (Eimeria tenella)感染鸡的盲肠、肝脏、脾脏和法氏囊组织中脂质过氧化物 (L PO)含量和超氧化物歧化酶 (SOD)、谷胱甘肽过氧化物酶 (GXH- Px)及过氧化氢酶 (CAT)活性 ,研究雏鸡感染球虫后机体内自由基生成和抗自由基反应。结果表明 ,雏鸡感染 E.tenella后 ,盲肠、肝脏和脾脏组织中的 L PO含量在感染后显著 (P<0 .0 5 )或极显著 (P<0 .0 1)升高 ,T- SOD、Mn- SOD、Cu、Zn- SOD、GSH- Px和 CAT活性水平不同程度下降。试验结果说明在 E.tenella感染后 ,鸡体内的氧自由基生成反应加剧 ,氧自由基参与了球虫的致病过程。     

不同禽流感病毒含量的新-流二联灭活疫苗免疫SPF鸡后抗体水平的监测

将未浓缩的新城疫抗原分别与未浓缩的、浓缩3倍、浓缩6倍的禽流感抗原混合,并制备成三组鸡新城疫、禽流感（H9N2 HP株）二联灭活疫苗（简称新-流二联灭活疫苗）,分别免疫21日龄SPF鸡,每羽0.3 mL,同时设置未免疫的空白对照组,免疫组与对照组均在免疫前及免疫后7、14、21、28、35 d进行采血,检测新城疫和禽流感抗体。结果发现,各免疫组在免后不同日龄的新城疫抗体基本一致,禽流感病毒抗原浓缩倍数越高（即禽流感病毒含量越高）的新-流二联灭活疫苗,免后14、21 d的抗体也越高;从免后21 d开始,各免疫组的禽流感抗体水平差异逐渐减小,免疫后禽流感抗体水平的高低可以反映该疫苗的免疫效果。试验结果表明,该疫苗可以通过浓缩提高抗原病毒含量的方法来提高免后早期抗体水平,取得良好的早期免疫效果。     

Distribution and Diversity of Salmonella Strains in Shipments of Hatchling Poultry,United States, 2013

Multistate outbreaks of salmonellosis associated with live poultry contact have been occurring with increasing frequency. In 2013, multistate outbreaks of salmonellosis were traced back to exposure to live poultry, some of which were purchased at a national chain of farm stores (Farm store chain Y). This study was conducted at 36 stores of Farm store chain Y and was concurrent with the timing of exposure for the human outbreaks of salmonellosis in 2013. We used environmental swabs of arriving shipment boxes of hatchling poultry and shipment tracking information to examine the distribution, diversity and anti‐microbial resistance of non‐typhoidal Salmonella (NTS) across farm stores and hatcheries. Isolates recovered from shipment boxes underwent serotyping, anti‐microbial resistance (AMR) testing and pulsed‐field gel electrophoresis (PFGE). Postal service tracking codes from the shipment boxes were used to determine the hatchery of origin. The PFGE patterns were compared with the PFGE patterns of NTS causing outbreaks of salmonellosis in 2013. A total of 219 hatchling boxes from 36 stores in 13 states were swabbed between 15 March 2013 and 18 April 2013. NTS were recovered from 59 (27%) of 219 hatchling boxes. Recovery was not significantly associated with species of hatchlings, number of birds in the shipment box, or the presence of dead, injured or sick birds. Four of the 23 PFGE patterns and 23 of 50 isolates were indistinguishable from strains causing human outbreaks in 2013. For serotypes associated with human illnesses, PFGE patterns most frequently recovered from shipment boxes were also more frequent causes of human illness. Boxes positive for the same PFGE pattern most frequently originated from the same mail‐order hatchery. Only one of 59 isolates was resistant to anti‐microbials used to treat Salmonella infections in people. This study provides critical information to address recurrent human outbreaks of salmonellosis associated with mail‐order hatchling poultry.     

Effect of breed of cattle on innate resistance to infection with <i>Babesia bovis, B bigemina and Anaplasma marginale

Objective To assess the innate resistance of naive Bos taurus, Bos taurus cross Bos indicus and Bos indicus cattle to virulent Babesia bovis, B bigemina and Anaplasma marginale parasites. Design Groups of 10, pure B indicus, fi B indicus cross,/B indicus cross and pure B taurus steers were infected with virulent B bovis, B bigemina and A marginale parasites. Procedure Sequential infections were carried out by intravenous inoculation of infected blood containing 1 times 10⁸ parasites of B bovis, followed by B bigemina and then A marginale. To assess resistance, measurements were made of parasitaemia, rectal temperature, packed cell volume and the number within a group requiring chemotherapy to control infection. There was a recovery period between each infection. Results Infection with B bovis showed that pure B indicus steers were significantly more resistant to B bovis infection than the other groups, with none of this group requiring treatment. There was no significant difference between fi B indicus cross and/B indicus cross with 30% and 20%, respectively, of steers in these groups requiring treatment. The pure B taurus steers were significantly more affected then those in the other three groups with 80% requiring treatment. Infections of B bigemina produced a mild response in comparison to that of B bovis and none of the steers required treatment. However, the pure B taurus group was significantly more affected than the other three groups for all other measurements. After the A marginale infection, B indicus steers were moderately affected with 50% requiring treatment, whereas 70% of the fi B indicus group, 80% of the /B indicus cross group and 100% of the pure B taurus group required treatment. Conclusions All breeds of cattle, ranging from pure B indicus to pure B taurus may be at risk of severe disease if exposed to virulent A marginale. The results confirm that pure B indicus cattle are relatively resistant to B bovis, but there could be a significant risk of severe mortalities if cross-bred herds are exposed to virulent infection.     

畜禽及环境中沙门氏菌的PCR快速检测与控制

本文运用PCR技术检测家畜及环境中的沙门氏菌,25种沙门氏菌均获得特异性扩增,3种非沙门氏菌检测结果为阴性。表明PCR技术具有快速简便、敏感高和特异性强等优点,值得在沙门氏菌的快速检测中推广应用。并简要介绍了沙门氏菌的相关控制措施。     

Plasma pepsinogen,inhibited larval development,and abomasal lesions in experimental infections of calves with Ostertagia ostertagi

Single doses of Ostertagia ostertagi, followed in 42 days by multiple increasing doses in calves, were monitored by fecal egg counts and plasma pepsinogen. The level of plasma pepsinogen increase was related to the increase in graded levels of inoculation and to the fecal egg counts. Plasma pepsinogen in uninfected controls remained below 1 IU/L. Plasma pepsinogen in all calves reached levels greater than 5 IU/L at 48 days of the extended inoculation period, although fecal egg counts remained low in the previously inoculated calves. The previously infected calves had a higher proportion of early fourth-stage larvae, while larger adult worm burdens were found in the previously uninfected calves. Early fourth-stage larvae were observed in extra-glandular sites, notably between the glandular epithelium and basement membrane or within the lamina propria. An immunological response of the host was suggested by the lymphoid cell infiltration in the mucosa. This host immune response may account for the greater larval inhibition exhibited by the previously infected calves.     

贵州省猪源大肠杆菌对四环素类药物的耐药性及耐药基因检测

为考察贵州省猪源大肠杆菌对四环素类抗菌药的耐药情况和耐药基因的流行情况,本试验采用微量肉汤稀释法测定783株分离于贵州省8个地区规模化养殖场的猪源大肠杆菌对6种四环素类药物的敏感性,并通过PCR方法检测其携带四环素耐药基因情况。结果显示,猪源大肠杆菌对土霉素、四环素、金霉素、多西环素、米诺环素和替加环素的耐药率分别为97.3%、97.6%、96.6%、89.3%、48.0%和34.2%;土霉素和四环素的耐药水平最高,其MIC₅₀分别为256 和128 μg/mL,而MIC₉₀均为512 μg/mL,四环素的耐药倍数高于土霉素;米诺环素和替加环素的耐药水平最低,其MIC₉₀分别为32和4 μg/mL,耐药倍数相同;耐药基因tetA、tetB、tetC、tetD和tetM的检出率分别为92.60%、41.50%、58.75%、58.62%和70.10%;耐药菌株中主要以tetA基因为主,且大多数以携带复合基因的形式存在;大肠杆菌对土霉素、四环素、金霉素、多西环素和米诺环素的耐药性分别与耐药基因tetB和tetM呈极显著相关（P<0.01）,tetD基因分别与对米诺环素和替加环素的耐药性呈极显著相关（P<0.01）。贵州省猪源大肠杆菌对四环素类药物的耐药情况十分普遍,尤其是四环素和土霉素,金霉素和强力霉素有高度耐药的趋势;外排泵是四环素类药物主要的耐药机制;在兽医临床上越来越严重的耐药情况与耐药基因的广泛存在有很大关系。     

狐肺炎大肠杆菌毒力基因和耐药基因检测及药物敏感性分析

为了解河北地区狐肺炎大肠杆菌毒力基因、耐药基因的分布及耐药性情况,本研究采用PCR技术对从狐肺炎病例分离的81株大肠杆菌的毒力基因和耐药基因进行检测,采用K-B法检测这些大肠杆菌对16种抗菌药物的敏感性,采用试管二倍稀释法检测中药对大肠杆菌的最小抑制浓度(MIC)和最小杀菌浓度(MBC),采用腹腔注射法检测81株大肠杆菌对小白鼠的致病性。结果显示:此次共分离到81株大肠杆菌,且对小白鼠具有较强致病性;毒力基因irp2、fyuA、Ler、eaeA检出率分别为50.6%、56.8%、9.9%、23.5%;耐药基因blaTEM、blaOXA1、sul1、sul2、blaTEM1、blaSHV-1、blaCTX-M、strA-strB、aadA1、blaPSE1、sul3、tetA、tetB、tetC、tetD、ermF检出率分别为84.0%、69.1%、43.2%、38.3%、28.4%、4.9%、24.7%、29.6%、16.1%、9.9%、11.1%、9.9%、18.5%、8.6%、13.6%、3.7%,未检出耐药基因ermB、ermC;81株大肠杆菌均对磺胺间甲氧嘧啶、林可霉素等耐药;诃子、乌梅、五味子对81株大肠杆菌具有较好的体外抑菌作用,抑菌圈直径在20 mm～36 mm,MIC和MBC在7.81 mg/mL～62.50 mg/mL。本研究调查了河北地区狐肺炎大肠杆菌的流行情况,对狐肺炎病的防治具有重要的指导意义,更具公共卫生学意义。     

宁夏部分地区腹泻羔羊源大肠埃希菌的分离鉴定与药敏试验

为掌握宁夏地区羔羊腹泻源大肠埃希菌的耐药性,采集腹泻羔羊肛拭子90份,开展大肠埃希菌的分离鉴定及药敏试验。结果分离到78株大肠埃希菌,对头孢类、氨基糖苷类、喹诺酮类、氟苯尼考、青霉素类、四环素等抗菌药物有较高的敏感性,对万古霉素、复方新诺明、克林霉素耐药(耐药率78.21%~94.87%)。结果表明,宁夏地区羔羊腹泻源大肠埃希菌对抗菌药物保持较高的敏感率,耐药现象并不严重。     

Clinical features associated with seroconversion to Anaplasma marginale, Babesia bigemina and Theileria parva infections in African cattle under natural tick challenge

A longitudinal study was conducted in Southeast Uganda for 14 months on 640 Zebu cattle kept under natural tick challenge, with a view to identifying clinical features for prediction of seroconversion to Anaplasma marginale, Babesia bigemina and Theileria parva infections. Physical examination, condition scoring and tick counts were undertaken on all cattle every 4 weeks. In addition, 5300 sera were collected and analysed for antibodies against A. marginale, B. bigemina and T. parva infections using the enzyme-linked immunosorbent assay (ELISA). The major clinical features compiled included weight loss, fever (rectal temperature), anaemia (packed cell volume), pallor of mucous membranes, lymph node enlargement, staring coat, diarrhoea and lacrymation. The risk factors included tick challenge at village level, sex, age, Rhipicephalus spp. density and Boophilus spp. density on individual animals. Using a binary logistic regression model, the clinical features and risk factors were analysed. The results suggest that increasing rectal temperature was associated with increased probability for seroconversion to A. marginale, while high level of Rhipicephalus spp. density and increasing packed cell volume (PCV) were significantly associated with reduced probability of seroconversion. Although statistically significant, none of the factors had large effects, with odds ratios (OR) of 0.87, 1.15 and 0.98 for Rhipicephalus spp. density, rectal temperature and PCV, respectively. For B. bigemina infection, a high level of Boophilus spp. density, anaemia and staring coat were significantly associated with increased probability of seroconversion (OR 1.50, 1.78, 1.37, respectively). Presence of lacrymation and old age were associated with reduced probability of seroconversion (OR 0.52, 0.86 respectively). For T. parva infection, lymph node enlargement (OR 1.30) was associated with increased probability of seroconversion, while high Rhipicephalus spp. density and increasing packed cell volume (PCV) were associated with reduced probability of seroconversion (OR 0.68 and 0.98, respectively). In conclusion, presence and intensity of the respective tick vectors for tick-borne diseases, age and clinical features such as anaemia, fever, staring coat, lymph node enlargement and lacrymation are indicators for seroconversion to A. marginale, B. bigemina and T. parva infections in cattle. These indicators for seroconversion could be exploited in the development of decision support tools for clinical diagnosis of tick-borne diseases.     

鸭源巴氏杆菌的分离鉴定及药敏试验

本试验为确定江西新余地区鸭发病的病原,对3个鸭场的病死鸭进行细菌分离,并通过形态特征、培养特性和生化试验对分离菌进行鉴定。从病死鸭的肝脏和心脏中分离到5株鸭多杀性巴氏杆菌,且分离菌对试验鸭和小白鼠有较强的致病性。药敏试验结果表明,分离的5株菌对左氧氟沙星、卡那霉素、头孢西丁、先锋必和大观霉素较为敏感。