First report of Calonectria cerciana causing leaf blight of Eucalyptus in northern India

Eucalyptus spp. and their hybrids are frequently cloned and mass planted across farmland tracts and commercial plantations in northern India. It is a viable feeder species to the paper and pulp industries in this region. In 2018 and 2019, during field surveys conducted in northern India, a serious leaf blight disease was frequently observed in E. tereticornis plantations. Isolation from the blighted leaf samples consistently yielded fungal isolates having Calonectria‐like morphology. Morphological features coupled with sequence analysis of partial β‐tubulin (TUB2) and partial translation elongation factor‐alpha (TEF1) gene regions of two fungal isolates confirmed the species as Ca. cerciana. In detached leaf assays and glasshouse inoculation experiments, both isolates produced symptoms similar to those observed on the naturally infected leaves. Koch's postulates were fulfilled by re‐isolating Ca. cerciana from the inoculated leaves. This work is the first to confirm that Ca. cerciana is associated with a serious leaf blight disease of Eucalyptus in northern India and is an important addition to the taxonomy of Calonectria fungi in India.     

In vitro effects of Musa x paradisiaca extracts on four developmental stages of Haemonchus contortus

This study was carried out to evaluate the in vitro effect of Musa x paradisiaca stem and leaf against the parasitic nematode of small ruminants Haemonchus contortus. Three extracts (aqueous, methanolic and/or dichloromethane) of Musa x paradisiaca stem and leaf were tested in vitro on four developmental stages of H. contortus using egg hatch assay (EHA), larval development assay (LDA), L3 migration inhibition assay (LMI) and adult worm motility assay (AWM). The highly significant (P < 0.0001) ability to stop larval development (inhibition >67% for each extract) and the negative effect of the dichloromethane extract of leaf on adult worm motility (43% of inhibition of motility after 24 h of incubation) compared to the negative controls, suggest anthelmintic properties of Musa x paradisiaca stem and leaf against H. contortus. The active principles responsible for the activity could be secondary metabolites such as terpenoid and flavonoid compounds present in the leaf and stem of the plant.     

A new Brucella canis species-specific PCR assay for the diagnosis of canine brucellosis

Brucellosis is a zoonotic disease that is transmitted from animals to humans, and the development of a rapid, accurate, and widely available identification method is essential for diagnosing this disease. In this study, we developed a new Brucella canis species-specific (BcSS) PCR assay and evaluated its specificity and sensitivity. A specific PCR primer set was designed based on the BCAN_B0548-0549 region in chromosome II of B. canis. The PCR detection for B. canis included amplification of a 300-bp product that is, not found on other Brucella species or, genetically or serologically related bacteria. The detection limit of BcSS-PCR assay was 6 pg/μl by DNA dilution, or 3 × 10³ colony-forming units (CFU) in the buffy coats separated from whole blood experimentally inoculated with B. canis. Using the buffy coat in this PCR assay resulted in approximately 100-times higher sensitivity for B. canis as compared to detect directly from whole blood. This is the first report of a species-specific PCR assay to detect B. canis, and the new assay will provide a valuable tool for the diagnosis of B. canis infection.     

检测水貂出血性肺炎绿脓杆菌的比色LAMP法的建立与应用

为了快速有效检测水貂出血性肺炎病原绿脓杆菌,本研究结合金属指示剂HNB特性建立了快速检测绿脓杆菌比色LAMP法。根据绿脓杆菌外毒素A基因设计引物,建立了检测LAMP法;并且对该方法进行了特异性、灵敏性分析;同时进行了初步应用。结果显示,该方法特异性强,灵敏性好,可检测167CFU/mL的菌体,对病貂肺脏检出率高。结果表明,该比色LAMP法可以有效地检测水貂出血性肺炎绿脓杆菌。     

城市废水暴露对食蚊鱼肝脏EROD酶活性的影响

采用动力学酶标荧光法,检测了东莞市数所污水处理厂、制药厂和电子厂废水对食蚊鱼(Gambusia affinis)肝组织中7-ethoxyresorufin o-deethylase(EROD)酶活性的影响,评价了运用EROD酶活性监测水环境污染物的生物效应的可行性。结果显示,食蚊鱼分别暴露于经稀释为20%,40%,60%,80%不同梯度的废水液72 h后,肝脏EROD酶的活性分别与受试城市污水处理厂、制药厂和电子厂的废水之间存在剂量效应关系,EROD酶活性随污水浓度的增加而提高。电子厂废水的最大诱导倍数与对照组的比值可达到5.26,这表明其水体中存在的有机污染物较多,污水处理厂次之,制药厂的出水中污染物最少。研究表明,食蚊鱼肝组织EROD酶活性可以作为监测城市废水污染的理想生物标记物,后续的研究工作应使之标准化。     

乙纶发光网线在张网拖网上的初步应用

试验结果表明，发光网线应用于被动性张网渔具时，增产效果不明显，而且还必须考虑资源保护和解决光源激发问题。在拖网渔具上应用时，能提高渔获效率，夜间生产效果高于白天，而且调整方便。     

The fate of chlortetracycline residues in a simulated chicken–fish integrated farming systems

Two experiments were conducted at the Asian Institute of Technology, Pathumthani, Thailand to investigate the fate of chlortetracycline (CTC) residue in chicken manure and its effect on integrated chicken–fish farming system. During the first experiment, broiler chickens were raised and CTC residues in their manure were analysed. Chicken fed diets containing 0, 50, 200 and 800 CTC mg kg^?1 had CTC residue levels of 0, 0.9, 3.8 and 6.5 CTC ng g^?1. Once the diet containing CTC was withdrawn, CTC in the manure dropped to negligible amounts (0, 0, 0.2 and 0.5 CTC ng g^?1) within 1 day. Integrated chicken–fish farming systems were simulated during the second experiment to determine the fate of antibiotic residues in chicken manure in aquaculture environment. Chickens were fed a CTC‐free diet and a feed containing CTC at 200 mg kg^?1. Ten 4 m³ square concrete tanks (2 × 2 × 1 m) were used for the experiment. Five tanks were fertilized with CTC‐contaminated manure and the remaining five tanks were fertilized with CTC‐free manure at a rate of 100 kg dry matter ha^?1 day^?1. Sex‐reversed Nile tilapia (Oreochromis niloticus) was stocked at 12 fish tank^?1 on the 14th day after chicken manure application. The immuno‐radio microbial receptor assay (Charm II test) revealed that edible fish muscle, fish intestinal tract and sediment were contaminated by CTC at rates of 7.21, 22.104 and 1.788 ng g^?1, respectively, after 45 days. Chlortetracycline was detected on day 20 in the water column and gradually increased from 0.26 to 12.13 ng g^?1. Chlortetracycline residues were not detected in fish or the aquatic environment of the CTC‐free treatment. The results demonstrate the potential for antibiotic residue accumulation in fish and aquatic environment when CTC‐contaminated chicken manure is used for pond fertilization.     

Analysis of exotoxins produced by atypical isolates of Aeromonas salmonicida,by enzymatic and serological methods

In this study, exotoxins produced by 62 Aeromonas salmonicida strains and the bacterium Haemophilus piscium were analysed. Enzymatic assays, zymograms and serological detection were used to monitor secretion by bacterial strains of the previously described exotoxins P1, GCAT and AsaP1 and also the extracellular P2 metallo-gelatinase and a serine caseinase, which is different from the P1 protease and has not yet been characterized. Based on the results, the strains were divided into five groups. One comprised the type strains for A. salmonicida ssp. masoucida, H. piscium and 36% of the atypical isolates, and another, a type strain for A. salmonicida ssp. smithia together with 14% of the atypical isolates. A second type strain of A. salmonicida ssp. smithia was grouped with 8% of the atypical isolates. The largest group contained the type strains for A. salmonicida ssp. achromogenes and 38% of the atypical isolates. The type strains for A. salmonicida ssp. salmonicida were in the last group with all the four typical strains and 4% of the atypical isolates. The combination of zymogram and serological detection used is recommended as the most reliable method for characterizing A. salmonicida strains according to their exotoxin secretion.     

口蹄疫灭活疫苗蛋白质含量测定操作程序的确立

为优化用于口蹄疫灭活疫苗蛋白质含量测定的改良Lowry法,进而确立口蹄疫灭活疫苗蛋白质含量测定的操作程序,探索了有机溶剂破乳剂、酚红以及丙酮沉淀对测定结果影响。结果表明：样品中含酚红和有机溶剂均导致测定值较标准值高;有机溶剂破乳后,水相样经过丙酮沉淀测定值较标准值低;丙酮直接沉淀疫苗后测定蛋白质值与标准值符合度最高,丙酮沉淀回收率随蛋白浓度升高而升高,回收率在90％～100％之间。试验首次确立了改良Lowry法检测口蹄疫灭活疫苗中蛋白质含量的操作程序为丙酮直接沉淀疫苗后测定蛋白质浓度。并成功应用于口蹄疫灭活疫苗蛋白质含量的测定。     

重庆市禽白血病及禽网状内皮组织增生症的血清流行病学调查

禽白血病和禽网状内皮组织增生症均为禽的肿瘤性免疫抑制疾病,是危害养鸡业的两种非常重要的病毒性传染病。为调查重庆市肉鸡中禽白血病及禽网状内皮组织增生症的流行情况,在北碚区、涪陵区、开县、垫江县、潼南县五个区（县）的8个活禽交易市场采集260份血清样本,采用酶联免疫吸附试验检测了所有血清中禽白血病病毒（ALV）和禽网状内皮组织增生病毒（REV）的抗体。检测结果显示：禽白血病病毒和禽网状内皮组织增生病毒抗体阳性率分别为16.5%（43/260）和5%（13/260）,双抗体阳性率为4.61%（12/260）。与全国其他地区相比,重庆市肉鸡中这两种病原的感染率相对要低,但仍应重视这两种疾病的防控,以控制病原的进一步传播。