Improving the specificity of immunodiagnosis for porcine brucellosis

This report describes the use of cell mediated immunity to improve specificity of current diagnosis for Brucella suis. Diagnosis is problematic due to cross reactions that lead to false positive serological reactions (FPSR) in the standard diagnostic tests. A common cause of this cross reactivity is infection with the organism Yersinia enterocolitica O:9. Gottingen™ mini-pigs were experimentally infected with B. suis biovar I field strain or Y. enterocolitica serotype O:9 biotype 3. Infection was followed for 70 days. During this time whole blood stimulation assays were set up using Brucella specific antigen. IFNγ was measured in the supernatants (SN) from these assays by ELISA. Concurrent standard serological tests were carried out. The results indicate that the IFNγ assay is specifically able to distinguish Y. enterocolitica O:9 infection from a B. suis infection in experimentally infected mini-pigs. These results represent an improvement in diagnostic specificity compared to currently used serological tests. Thus suggesting that in a surveillance setting this test could be applied as a confirmatory test in the face of FPSR. The authors are British Civil Servants and as such their work is subject to British Crown Copyright. This means the exclusive copyright for the article cannot be transferred.     

Prevalence and Aetiology of Mastitis in Cows from Two Major Ethiopian Dairies

The study was undertaken to determine the aetiology and prevalence of mastitis in hand-milked cows (n = 186) in two major Ethiopian dairies. The California Mastitis Test and culturing for bacteria revealed that 21.5% of the cows were clinically infected and 38.2% had subclinical mastitis. Most mastitis pathogens isolated from milk samples testing positive by the California Mastitis Test were Gram-positive cocci. Staphylococci constituted 57% of the isolates, of which the predominant cause of bovine mastitis was Staphylococcus aureus (40.5%). Other mastitis pathogens isolated include streptococci (16.5%), coliforms (9%) and corynebacteria (5%). Retrospective analysis of farm records indicated that mastitis was the second most important cause of culling and accounted for 27% of the cows removed from these two dairies.     

Efficacy of Sarafloxacin in Broilers after Experimental Infection with Escherichia coli

Infections of chickens with Escherichia coli serotypeO78 can be treated with the antibiotic sarafloxacin. Three experiments were conducted on the administration of this drug to chickens that had been experimentally infected with E. coli. The birds were monitored for 10 days after infection for their average daily gain (ADG) and feed conversion ratio (FCR), and the post-mortem pathology was assessed. In the first experiment, sarafloxacin (20 mg/L, equivalent to 5 mg/kg live weight per day), given in the drinking water for 3 days after infection, led to a reduction in the mortality from 75% to 27%, but the ADG of the treated birds was still less than that of the uninfected controls. In the second experiment, when the sarafloxacin was administered at the same dose in the water but over only 2 h, there was also a considerable reduction in mortality, and the ADG and the FCR also improved significantly. In the third experiment, the dose dependence of the drug was tested. The birds were given 5 and 10 mg/kg per day sarafloxacin in each group, starting within 2 h after infection. This rapid administration of the drug completely prevented mortality, while the ADG and FCR were similar to those of the uninfected controls.     

Inhibitory effects of a monoclonal antibody (MAb-001) on in vitro oxygen consumption and multiplication of the pathogenic haemoflagellate, Cryptobia salmositica Katz

A monoclonal antibody (MAb-001), against a surface glycoprotein on Cryptobia salmositica inhibited the multiplication and oxygen consumption of both virulent and avirulent strains of the parasite. The classical cysteine proteinase inhibitor (E-64) and a cysteine proteinase activator (EDTA) affected the in vitro multiplication of C. salmositica . Concentrations of E-64 higher than 10 μ M reduced the multiplication of C. salmositica while 5 m M of EDTA enhanced its multiplication. We propose that the cysteine proteinase is an important metabolic enzyme in C. salmositica and that binding of MAb-001 to the enzyme inhibited parasite multiplication and reduced oxygen consumption.     

Demonstration of alternative feeds for the Pacific white shrimp, Litopenaeus vannamei, reared in low salinity waters of west Alabama

The replacement of marine proteins with vegetable proteins or terrestrial animal byproducts in aquaculture diets has been gaining momentum. This study examines the viability of replacing fish meal in shrimp production diets with alternative protein sources (combinations of vegetable proteins) in inland low salinity waters of west Alabama. The test diets were formulated to contain 36% protein and 8% lipid. The basal diet contained 10% fish meal. The fish meal was then replaced (on a weight to weight basis) with poultry meal (PM), pea meal or distiller's dried grain with solubles (DDGS). Two separate experiments (laboratory trial and farm trial) were devised to test the efficacy of the diets for Litopenaeus vannamei reared in low salinity waters. The laboratory trial was conducted at the E. W. Shell Fisheries Research Station in Auburn, Alabama, USA, whereas the farm trial examined the same diets in an outdoor flow-through system at a low salinity shrimp farm in west Alabama. Results indicate no significant differences in shrimp growth, weight gain, survival or feed conversion ratio among dietary treatments, suggesting that PM, pea meal and DDGS can serve as alternatives to fish meal as a protein source for shrimp reared in low salinity environments.     

High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.     

Influence of dietary fat level and whole-body adiposity on voluntary energy intake by juvenile rainbow trout Oncorhynchus mykiss (Walbaum) under self-feeding conditions

The influence of dietary fat level and whole‐body adiposity on voluntary energy intake of juvenile rainbow trout Oncorhynchus mykiss (Walbaum) was examined using self‐feeders. Groups of lean fish [crude fat (CF) = 7%] and fat fish (CF = 11%), pretreated with a commercial diet with or without supplemental pollock oil, were self‐fed one of three fat level diets (CF = 8%, 13.5% and 19%) for 48 days at 17 °C. Final body weight (BW) and total digestible energy (DE) intake (kJ per fish) were positively affected by the initial BW. Relative to the initial BW, however, fat fish consumed less DE than lean fish. Although the effect of dietary fat level was not significant, percentage weight gain and daily DE intake per BW (kJ kg^?1 BW day^?1) of fat fish were significantly lower than those of lean fish (ancova with initial BW as a covariate, P < 0.05). Energy digestibility, feed efficiency and protein retention were improved with the dietary fat level; however, there was no difference resulting from body fat level. The whole‐body fat levels at the end of the experiment increased with the dietary fat level. Between groups self‐fed the same diet, fat levels of the initially fat fish were still higher than those of the lean fish. The results of the present medium‐term study suggest that rainbow trout adjust DE intake from diets with fat levels ranging from 8% to 19%. Although body fat level affects neither energy digestibility nor protein utilization, a high body fat level may reduce DE intake and consequently depress growth.     

Effect of cryoprotectants, extenders and freezing rates on the fertilization rate of frozen striped catfish, Pangasius hypophthalmus (Sauvage), sperm

The effects of four cryoprotectants (methanol, MeOH; dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; and ethylene glycol, EG), three extenders (calcium‐free Hanks' balanced salt solution, C‐F HBSS, Hanks' balanced salt solution, HBSS and sodium chloride, NaCl) and two different freezing procedures (one‐ and two‐step) on the cryopreservation of striped catfish (Pangasius hypophthalmus (Sauvage)) sperm were investigated. Sperm were frozen using a controlled‐rate freezer in 250 μL straws and stored for 2 weeks in a liquid nitrogen (LN₂) container. They were then airthawed at room temperature, and fertilization, motility and viability were assessed. The highest fertilization rate of 41% (81% of control) was achieved with the combination of 12% DMSO and 0.9% NaCl using a one‐step freezing procedure (10°C min^?1). Also, DMA resulted in a higher fertilization rate (30% or 51% of the control) than MeOH (18% or 38% of the control) or EG (8% or 12% of the control). In addition, the three extenders used did not affect fertilization rates after cryopreservation with each cryoprotectant. There were no significant differences among the three cryoprotectant concentrations and between the one‐ and two‐step freezing procedures. However, fertilization rates of cryopreserved sperm were significantly lower than the controls (P<0.05). The results of this study indicate that high fertilization rates of striped catfish eggs can be achieved using cryopreserved sperm when frozen at 10°C min^?1 in DMSO or DMA with either 0.9% NaCl or C‐F HBSS.     

Migratory history of the giant mottled eel (Anguilla marmorata) in the Bonin Islands of Japan

Chino N, Arai T. Migratory history of the giant mottled eel ( Anguilla marmorata ) in the Bonin Islands of Japan.
Ecology of Freshwater Fish 2010: 19: 19–25. © 2009 John Wiley & Sons A/S
Abstract –  In order to elucidate the migratory history and habitat use of the giant mottled eel Anguilla marmorata, in the Bonin Islands of Japan, strontium (Sr) and calcium (Ca) concentrations in otoliths were examined by X-ray electron microprobe analysis. The X-ray intensity map and line analyses showed three fluctuation patterns: (i) freshwater residence, (ii) continuous residence in brackish water, and (iii) residence in freshwater after recruitment, while returning to brackish water. Based on these results and previous studies, the migratory behaviours of A. marmorata appear to differ in each habitat in response to inter- and intra-specific competition. The results suggest that A. marmorata has a flexible pattern of migration, with an ability to adapt to various habitats and salinities.