Transmission of Cyprinid herpesvirus-3 (CyHV-3) from goldfish to naïve common carp by cohabitation

Cyprinid herpesvirus-3 (CyHV-3) has spread worldwide and has had a major impact on koi and common carp production. Previous studies on the host range of the CyHV-3 found that fish species other than koi and common carp are fully resistant to natural virus exposure. Recently, CyHV-3 was detected in goldfish (Carassius auratus auratus) that were in contact with CyHV-3 infected koi. In the present study, a specific RT-PCR product was amplified from the viral thymidine kinase gene in gills, intestine and brain tissues of CyHV-3 infected goldfish. This implied that CyHV-3 replicated in these goldfish. Also, in the presence of a stress factor such as temperature fluctuation, the CyHV-3 infected goldfish transmitted the virus to cohabitated naïve SPF common carp. CyHV-3 DNA was detected in the cohabitated naïve carp tissues by PCR. The results of this study demonstrate that goldfish is a carrier for CyHV-3, permit virus propagation, and disseminate the virus to susceptible carp causing the disease.     

The use of a semiochemical bait to enhance exposure of Amblyomma variegatum (Acari: Ixodidae) to Metarhizium anisopliae (Ascomycota: Hypocreales)

Experiments were conducted to explore the use of a semiochemical bait to enhance exposure of Amblyomma variegatum Fabricius (Acari: Ixodidae) to different formulations of the entomopathogenic fungus Metarhizium anisopliae (Metsch.) Sorok. (Ascomycota: Hypocreales). Initially, the relative efficacies of attraction-aggregation-attachment pheromone (AAAP), made up of o-nitrophenol, methyl salicylate and nonanoic acid in the ratio 2:1:8, 1-octen-3-ol and butyric acid, were evaluated in an olfactometer. Only AAAP and 1-octen-3-ol were found to elicit attractive responses to the tick. Simultaneous release of 1-octen-3-ol and AAAP together with CO₂ from a trap in semifield plots attracted up to 94.0 ± 6% of adult ticks from a distance of 6 m, and up to 24.0 ± 5.1% from 8 m. Formulations of M. anisopliae (dry powder, oil, and emulsifiable) applied within the trap baited with AAAP, 1-octen-3-ol and CO₂ resulted in high levels of contamination of the ticks attracted to the traps. However, 48 h after autoinoculation, 89.1 and 33.3% of conidia were lost in dry powder and oil formulations, respectively. Emulsifiable formulation showed least loss of propagules (17.1%). Samples of ticks attracted to the baited traps were transferred to plastic basins containing grass and maintained for 5 weeks. The experiment was conducted in rainy and dry seasons. Emulsifiable formulation gave the highest relative tick reduction in both seasons: 54.7 and 46.5% in rainy and dry seasons, respectively, followed by oil formulation (32.0 and 23.8%) and powder formulation (38.0 and 24.4%).     

Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD)

Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout.     

Resistance patterns and detection of aac(3)-IV gene in apramycin-resistant Escherichia coli isolated from farm animals and farm workers in northeastern of China

The aminoglycoside apramycin has been used widely in animal production in China since 1999. This study was aimed to investigate the resistance pattern of apramycin-resistant Escherichia coli isolated from farm animals and farm workers in northeastern of China during 2004–2007 and to determine whether resistance to apramycin was mediated by plasmid containing the aac(3)-IV gene and the mode for the transfer of genetic information between bacteria of farm animals and farm workers. Thirty six E. coli isolates of swine, chicken, and human origins, chosen randomly from 318 apramycin-resistant E. coli isolates of six farms in northeastern of China during 2004–2007, were multi-resistant and carried the aac(3)-IV gene encoding resistance to apramycin. Conjugation experiments demonstrated that in all 36 cases, the gene encoding resistance to apramycin was borne on a mobilisable plasmid. Homology analysis of the cloned aac(3)-IV gene with the sequence (accession no. X01385) in GenBank showed 99.3% identity at a nucleotide level, but only with a deletion of guanosine in position 813 of the gene in all 36 cases. The results indicted that resistance to apramycin in these isolates was closely related to aac(3)-IV gene. Therefore, the multi-resistance of E. coli could complicate therapeutic practices for enteric infections in both farm animals and human.     

The supplementary value of different parts of enset (Ensete ventricosum) to sheep fed wheat straw and Desmodium intortum hay

A study which included growth and digestibility experiments was conducted to evaluate the feeding value of a) enset pseudostem, b) enset corm or c) enset mixture (containing equal amounts of pseudostem, corm and enset leaf on a DM basis) as a supplement to a basal diet of wheat straw fed ad libitum and restricted amounts of Desmodium intortum (Desmodium) hay to sheep. During the 71-day growth experiment, 18 one-year-old male sheep of indigenous breed with mean body weight (BW) of 20.2 (± 0.43) kg were randomly assigned to the three treatments. In the digestibility experiment, three male sheep with mean BW of 19.5 (± 1.06) kg were assigned to each treatment. Sheep supplemented with enset mixture consumed more (P < 0.001) wheat straw DM (335 g/day) than those supplemented with pseudostem (295 g/day). No differences were found in total dry matter (DM) and organic matter (OM) intake between diets supplemented with pseudostem, corm and enset mixture. Total nitrogen (N) intake (g/day) was higher (P = 0.0073) in sheep fed the diet with enset mixture (12.2) followed by corm (11.3) and pseudostem (10.7). The apparent digestibility of DM was higher (P < 0.001) in pseudostem (0.53) followed by corm (0.46) and enset mixture (0.41) supplemented sheep. Organic matter, neutral detergent fiber and N digestibility were similar among treatments. The amount of N lost in urine and faeces and the N-retention was similar among treatments. There were no differences in gain/day in sheep fed diets with pseudostem (29.3 g), corm (27.0 g) and enset mixture (36.5 g). It is concluded that supplementation of a wheat straw/Desmodium diet with enset fractions could be used equally at least to maintain body weight during the dry season.     

Quantification of water buffalo (Bubalus bubalis) cytokine expression in response to inactivated foot-and-mouth disease (FMD) vaccine

This study describes the quantification of cytokine expression of vaccinated water buffaloes with FMD inactivated vaccine. Using real-time PCR quantification assay, expression of Th1 (IL-2, IL-12p40, IFNγ); Th2 (IL-4, IL-10) and inflammatory (IL-6, TNFα) cytokines were quantified weekly for the entire three-week duration of the experiment. It was noted that IFNγ, IL-10 and TNFα had peaked on week three post-vaccination while the remaining cytokines peaked on the second week and decreased by the third week. The counteraction between IFNγ and IL-4 was noted as well as the possible suppressive action of IL-10 to that of IL-2 and IL-12, which is a common phenomenon between Th1 and Th2 cytokines. Synergy between TNFa and IL-6 was also observed. These findings suggest that within the immune system of water buffalo there is a dynamic cell-mediated and humoral interaction in response to immunogen. This assessment of the cytokine expressions is vital for the study of water buffalo disease progression and concurring protective immune responses.     

Sarcocystis neurona infections in raccoons (Procyon lotor): evidence for natural infection with sarcocysts, transmission of infection to opossums (Didelphis virginiana), and experimental induction of neurologic disease in raccoons

Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses in the Americas and Sarcocystis neurona is the most common etiologic agent. The distribution of S. neurona infections follows the geographical distributions of its definitive hosts, opossums (Didelphis virginiana, Didelphis albiventris). Recently, cats and skunks were reported as experimental and armadillos as natural intermediate hosts of S. neurona. In the present report, raccoons (Procyon lotor) were identified as a natural intermediate host of S. neurona. Two laboratory-raised opossums were found to shed S. neurona-like sporocysts after ingesting tongues of naturally-infected raccoons. Interferon-gamma gene knockout (KO) mice fed raccoon-opossum-derived sporocysts developed neurologic signs. S. neurona was identified immunohistochemically in tissues of KO mice fed sporocysts and the parasite was isolated in cell cultures inoculated with infected KO mouse tissues. The DNA obtained from the tongue of a naturally-infected raccoon, brains of KO mice that had neurological signs, and from the organisms recovered in cell cultures inoculated with brains of neurologic KO mice, corresponded to that of S. neurona. Two raccoons fed mature S. neurona sarcocysts did not shed sporocysts in their feces, indicating raccoons are not likely to be its definitive host. Two raccoons fed sporocysts from opossum feces developed clinical illness and S. neurona-associated encephalomyelitis was found in raccoons killed 14 and 22 days after feeding sporocysts; schizonts and merozoites were seen in encephalitic lesions.     

Pharmacokinetics of marbofloxacin after a single oral dose to loggerhead sea turtles (Caretta caretta)

The single-dose disposition kinetics of marbofloxacin (MBX) were determined in clinically healthy loggerhead sea turtles (n = 5) after oral (PO) administration of 2 mg kg⁻¹ bodyweight. Marbofloxacin plasma concentrations were determined by DAD–HPLC (LOD/LOQ 0.015/0.05 μg ml⁻¹). Data were subjected to non-compartmental analysis. Following PO administration, marbofloxacin achieved maximum plasma concentrations of 11.66 ± 2.53 mg L⁻¹ at 15.00 ± 3.00 h. The absence of general adverse reactions in the turtles of the study, and the favourable pharmacokinetic properties (long half-life and high maximum plasma concentration) of MBX administered PO at the single-dose of 2 mg kg⁻¹ suggest the possibility of its safe and effective clinical use in loggerhead sea turtles.     

The binding of soybean agglutinin (SBA) to the intestinal epithelium of Atlantic salmon, Salmo salar and Rainbow trout, Oncorhynchus mykiss, fed high levels of soybean meal

A lectin present in soya, soybean agglutinin (SBA), was identified in electrophoretic profiles and immunoblots of dehulled solvent-extracted soybean meal (DSSM), full-fat soybean meal (FFSM) and of aqueous extracts of feeds incorporating them in their formulation. A quantitative estimation was made of the proportion of SBA comprising the total protein in FFSM and a trial diet was prepared containing an amount of pure SBA similar to that in diets incorporating high levels of the whole soya product. Fish fed with this diet exhibited similar pathological disruption of the intestinal tract to that observed in fish given a diet with a high level of DSSM (60% of the diet). Furthermore, immuno-histochemistry revealed the binding of the SBA to the enterocytes lining the intestinal villi both of fish fed a diet incorporating pure SBA and those fed a diet containing a high-level of soya (60%). Our results suggest that SBA binds in vivo to the intestinal epithelium of fish and has a contributory role in pathological changes associated with fish feeds containing high levels of soybean proteins.     

Use of a Mycobacterial Cell Wall Extract (MCWE) in Susceptible Mares to Clear Experimentally Induced Endometritis With Streptococcus zooepidemicus

The ability of an immunomodulator, mycobacterial cell wall extract (MCWE), to clear uterine infection in susceptible mares after an experimental challenge withStreptococcus zooepidemicus was evaluated. Thirty mares susceptible to endometritis, based on the presence of uterine fluid during both diestrus and estrus, were selected from a herd of 896 and inoculated with a live culture of 5 × 10⁶ CFU of S. zooepidemicus on day 1 of estrus. Twenty-four hours later, mares were evaluated by ultrasonography, bacteriology, exfoliative cytology, and uterine biopsy to confirm infection. Forty-eight hours after inoculation, and on confirmation of uterine infection, mares were randomly assigned to one of four unbalanced experimental treatments to receive 1500 μg MCWE IU (n = 10) or IV (n = 10), or placebo IU (n = 5) or IV (n = 5). Mares were examined at ovulation and 7 days post-ovulation for uterine fluid via transrectal ultrasonography and for bacteriology, exfoliative cytology, and uterine biopsy. Efficacy was based on the ability of the mare to clear endometritis as determined by negative bacteriology and reduced numbers of polymorphonuclear cells (PMNs) on uterine biopsy. Because no statistical difference was detected between routes of administration on day 7 post-ovulation, the data sets were combined and re-analyzed to evaluate overall efficacy. Endometritis was observed in all placebo-treated mares 7 days post-ovulation, whereas treatment with MCWE resulted in the elimination of endometritis in 35% of the mares by the time of ovulation, and 70% of the mares by 7 days post-ovulation. Treatment with MCWE, compared with the placebo group, resulted in a significant decrease in the number of mares positive for endometritis at ovulation based on exfoliative cytology and bacteriology (P < .01) and at 7 days post-ovulation based on biopsy, exfoliative cytology, and bacteriology (P < .001). Results indicate that MCWE was an effective treatment for the elimination of endometritis caused by S. zooepidemicus in mares.     

Association between risk of seroconversion of sentinel cattle to bluetongue viruses and Culicoides species (Diptera: Ceratopogonidae) in Queensland, Australia

The association between risk of seroconversion of sentinel cattle to bluetongue viruses and the number of Culicoides brevitarsis Kieffer and C. wadai Kitaoka caught by light traps was investigated using survival analysis. Eight sentinel herds that seroconverted to bluetongue viruses between 1990 and 1994, and for which insect-trapping data were available, were selected for inclusion in the study. These herds were located at six sites along the eastern coast of Queensland, Australia, from approximately latitude 10 °South to 25 °South. C. brevitarsis was detected at all locations where sentinel herds were maintained, whereas C. wadai was detected at only two locations in northern Queensland where four sentinel herds were maintained during the study period. The mean number of C. brevitarsis and C. wadai caught per month was 230 and 21, respectively. A significant (P = 0.05) positive association was found between the risk of seroconversion of sentinel cattle to bluetongue viruses and the number of C. wadai caught in the same month.     

Use of refined potato (Solanum tuberosum L. cv. Gogu valley) protein as an alternative to antibiotics in weanling pigs

This study investigates the use of refined potato protein (RPP) obtained from the potato (Solanum tuberosum) tubers of “Gogu valley” for their antimicrobial properties and its effects on growth performance, apparent nutrient digestibility and microbial populations in feces and large intestine of weanling pigs. Pigs (n = 280; Landrace × Yorkshire × Duroc; average initial body weight of 5.96 ± 0.66 kg; 23 ± 3 d of age) were allotted on the basis of their body weight and sex to five dietary treatments (each treatment comprised of 4 replicate pens with 14 pigs in each) in a randomized complete block design. The dietary treatments were: PC (positive control; basal diet + 150 ppm apramycin sulfate and 10 ppm colistin sulfate), and RPP (basal diet added with 0, 200, 400 and 600 ppm RPP). The RPP showed in vitro antimicrobial activity and at the concentration of 150 ppm inhibited the growth of tested microbes (Staphylococcus aureus, Salmonella choleraesuis, Salmonella gallinarum and Escherichia coli). Pigs fed with antibiotic diet showed better growth performance and had lower populations of total bacteria, coliforms and Staphylococcus spp. in the feces and large intestine when compared with pigs fed RPP diets. Increasing the levels of RPP in diets linearly improved performance and reduced the populations of total bacteria, coliforms and Staphylococcus spp. in feces and contents of colon and rectum. The apparent fecal digestibility of DM, CP and apparent ileal digestibility of amino acids did not differ among pigs fed antibiotic and RPP diets. These results suggest that antibiotics were more effective in improving the performance of pigs, while feeding of RPP also improved the performance and reduced the harmful microbes being more effective at higher levels. Thus at higher levels RPP obtained from the potato tubers of Gogu valley can be a potential replacement of antibiotics in the feed of weanling pigs.     

Effect of varying ruminally degradable to ruminally undegradable protein ratio on nutrient intake, digestibility and N metabolism in Nili Ravi buffalo calves (Bubalus bubalis)

Four Nili-Ravi buffalo calves (100 ± 4 kg) were used in 4 × 4 Latin Square Design to evaluate the influence of varying ruminally degradable protein (RDP) to ruminally undegradable protein (RUP) ratio on dry matter intake (DMI), digestibility and nitrogen (N) metabolism. Four experimental diets A, B, C and D were formulated to contain RDP:RUP of 70:30, 65:35, 60:40 and 55:45, respectively. The calves were fed ad libitum. Dry matter intake by calves fed C diet was higher (P < 0.05) than those fed D diet and lower (P < 0.05) than calves fed A diet, however, it was similar to those fed B diet. There was a linear decrease (P < 0.01) in DMI with decreasing the RDP to RUP ratio. Similar trend was noticed in crude protein (CP) intake. Neutral detergent fiber (NDF) intake was significantly different across all treatment. The decrease in CP and NDF intake was due to decreasing trend of DMI. Dry matter (DM) digestibility in calves fed A and B diets was higher (P < 0.05) than those fed C and D diets. A linear decrease (P < 0.01) in DM digestibility was observed with decreasing the RDP to RUP ratio. Crude protein digestibility remained unaltered across all treatments. Neutral detergent fiber digestibility was higher in calves fed A and B diets than those fed C and D diets. Higher NDF digestibility in calves fed A and B diets was due to higher level of dietary RDP that might resulted in higher ruminal ammonia concentration which stimulate activity of cellulytic bacteria and ultimately increased NDF digestibility. The N retention (g/d) was similar among the calves fed B, C and D diets, however, it was higher (P < 0.05) than those fed A diet. Decreasing the RDP to RUP ratio resulted in linear increase (P < 0.01) in N retention. The N retention, as percent of N intake was significantly different across all treatments. Decreasing RDP to RUP ratio resulted in linear increase (P < 0.01) in N retention, as percent of N intake. A similar trend was noticed in N retention, as percentage of N digestion. Blood urea nitrogen (BUN) concentration in calves fed B diet was higher (P < 0.05) than those fed D diet and was lower (P < 0.05) than those fed A diet, however, it was not different from calves fed C diet. Decreasing dietary RDP to RUP ratio resulted in linear decrease (P < 0.05) in BUN concentrations. The decrease in BUN concentration was because of decreasing level of dietary RDP. The N retention can be increased by decreasing RDP to RUP ratio in the diet of growing buffalo calves and diet containing RDP to RUP ratio 55:45 is considered optimum regarding N retention in buffalo calves.     

Reproductive performance of farmed red deer (Cervus elaphus) in New Zealand: V: Mob and individual-hind risk factors associated with calf rearing to weaning

A 2-year longitudinal observational study of 15 red deer farms was carried out in New Zealand from March 1992. Both years combined, 3364 adult and 653 yearling hinds were diagnosed pregnant in June by ultrasound scanning and monitored for reproductive success up to weaning. At weaning, pre-rut at approximately 100 days after calving, udders were palpated or observed by the farmer and hinds were categorised as lactating (wet) indicating survival of calf, or not lactating (dry). Farm management practices during calving and early lactation were recorded. Analysis of risk factors associated with progeny survival was done using path analysis (separately for adult and yearling hinds). Adult hinds >3 years old at calving, conceiving before 1 May and having a body-condition score >2 in September had 2.8, 2.4 and 2.9 times significantly higher odds of rearing a calf to weaning than other hinds, respectively. Adult hinds had about 5 times lower odds of rearing a calf to weaning when they were mixed with mature stags during calving. When farmers visited calving paddocks and checked for calving problems, adult hinds had 1.7 times higher odds of rearing a calf to weaning. When the farmer entered calving paddocks at least every second time they observed the hinds, yearling hinds had a twice higher odds of rearing a calf up to weaning. Sunny weather and high maximum temperatures were positively and negatively associated, respectively, with the probability of an adult hind being in lactation at weaning.     

Assessment of enzyme linked immunosorbent assay based diagnostic kits (Toxokit-G and Toxokit-M) for the detection of IgG and IgM antibodies to Toxoplasma gondii in human serum

An enzyme linked immunosorbent assay (ELISA) using penicillinase was developed in the form of diagnostic kits (Toxokit-G and Toxokit-M) for the detection of IgG and IgM antibodies to Toxoplasma gondii. The performance of both the kits was compared with commercially available diagnostic kits, i.e. Enzygnost-Toxoplasmosis/IgG (Behring Co., Germany), TOXOTEK-G (Flow Lab., U.K.) and Toxoplasma IgM Microassay (Diamedix Corp., U.S.A.) by testing toxoplasma-suspected human serum samples. The results indicate a good reliability between these diagnostic kits. Toxokit-G has 86.66 and 96.05% sensitivity and specificity respectively. The main advantage of Toxokit-G is that the end result can be assessed visually without using sophisticated instruments. Toxokit-M has 100% sensitivity and specificity and test results were not affected by the presence of antitoxoplasma IgG antibodies, rheumatoid factor or antinuclear antibodies.     

The inclusion of a daisy plant (Chrysanthemum coronarium) in dairy sheep diet. 1: Effect on milk and cheese fatty acid composition with particular reference to C18:2 cis-9, trans-11

An experiment was carried out to evaluate the effect on milk and cheese fatty acid composition of the inclusion of Chrysanthemum coronarium L., (Asteracea) into dairy sheep diet. Plots sown either with a binary mixture consisting of Lolium rigidum Gaudin and Medicago polymorpha (LM) or a ternary mixture including the above species and C. coronarium were (LMC) grazed by groups of Sarda dairy ewes during May. The botanical composition of diet, as measured by n-alkane method, showed that 34% of LMC group diet consisted of Chrysanthemum flowers and leaves. Exposure of sheep to this non-conventional forage did not affect animal performance and milk composition (fat and casein, P > 0.05) but resulted in different milk and cheese fatty acid composition. In particular conjugated linoleic acid, vaccenic acid and linoleic acid were all higher in LMC than LM groups (P < 0.05). The probable main reason was the higher proportion of linoleic acid in Chrysanthemum forage and hence in LMC than LM diet.