Impact of concentrations of camphor on the in vitro mixed ruminal microorganism fermentation from goats selected for consumption of low and high levels of Juniperus spp.-2.

The microbial population in the gastrointestinal tract of ruminant animals aids in the utilization of forages with high levels of secondary plant compounds. Two divergent bloodlines of meat goats have been selected by screening fecal samples with near-infrared reflectance spectroscopy to assess the goat's consumption of high or low levels of Juniperus sp. leaves containing several monoterpenes, including camphor. The mechanism by which these goats can consume greater concentrations of Juniperus spp. leaves than their counterparts is unclear, and therefore, this study was designed to determine if differences existed between the ruminal microbial populations of the low and high juniper-consuming bloodlines (LJC vs. HJC) by analyzing their ruminal microbiota and fermentation end products. In the present study, concentrations (0.00, 0.5, 0.99, 1.97, or 5.91 mM) of camphor were added to mixed ruminal microorganism fermentation. Five LJC and five HJC goats were fed a juniper-free diet (n = 10), and five LJC and five HJC goats (n = 10) were fed a diet that contained 30% fresh Juniperus ashei leaves for 21 d prior to ruminal fluid collection. In vitro fermentations used LJC and HJC, ruminal fluid inoculum added (33% v/v) to anoxic media in sealed Balch tubes. Camphor increased (P < 0.05) total short-chain fatty acid (SCFA) concentrations for all but one experimental group. Between the main dietary and bloodline goat effects, the diet was significant for all SCFA results except butyrate. In contrast, bloodline was only significant for acetate and butyrate molar proportions. Rumen fluid from juniper-free-fed goats exhibited greater concentrations of Ruminococcaceae, whereas juniper-fed goats contained more Coriobacteriaceae. Results demonstrated that mixed ruminal microorganisms fermentations from HJC goats did not produce greater concentrations of SCFAs or have the ability to degrade camphor at a higher rate than did that from LJC goats. Results suggest that camphor tolerance from J. ashei, was related to hepatic catabolic mechanisms instead of ruminal microbial degradation; however, further in vivo work is warranted.

Impact of camphor on the in vitro mixed ruminal microorganism fermentation from goats selected for consumption of low and high levels of Juniperus spp. 1.

Encroaching plant species in West Texas continues to significantly reduce livestock production capacity and cause an imbalance between plant and livestock ecology. Juniperus ashei and Juniperus pinchotii are encroaching species rarely used by browsing animals, mainly due to phytochemical defenses. Juniperus spp. contain large concentrations of monoterpenes in their essential oil profiles to deter herbivory. Since 2003, two divergent bloodlines of meat goats have been selected to consume low or high (LJC or HJC) amounts of juniper foliage, through screening fecal samples with near-infrared reflectance spectroscopy. However, it remains unclear whether HJC goats expressed a physiological ability to consume a greater amount of juniper or if they were colonized by a ruminal microbial population that could detoxify juniper phytochemicals. Therefore, this study aimed to investigate the impact of 0.00 and 1.97 mM of camphor on the mixed ruminal microorganism fermentation after 0, 1, 2, 4, 12, and 24 h of incubation. Five LJC and five HJC goats were fed a juniper-free diet (n = 10) and five LJC and five HJC goats (n = 10), were fed a diet containing 30% fresh J. ashei leaves for 21 days prior to ruminal fluid collection. In vitro fermentations used LJC and HJC, ruminal fluid inoculum was added (33% v/v) to anoxic media in sealed Balch tubes. Total short-chain fatty acid (SCFA) production and acetate to propionate ratio were increased (P < 0.05), but there was no effect on pH (6.56 ± 0.09). Goats that received the juniper-free diet had higher (P < 0.05) SCFA production than juniper-containing diets. There was no consistent difference in LJC and HJC microbial fermentation end products caused by the addition of 1.97 mM of camphor, and goats receiving a juniper-free diet consistently generated more SCFAs in the presence of 1.97 mM of camphor. Furthermore, bloodline differences in juniper consumption were likely related to physiological adaption capacities within the animal and not a ruminal microbial detoxification advantage.

Antimicrobial Susceptibility of Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli Isolated from Mastitic Dairy Cattle in Ukraine.

Bovine mastitis is the predominant cause for antimicrobial use on dairy farms and is a major source of economic losses in the dairy industry. In this study, the antimicrobial susceptibility profiles of common mastitis-causing pathogens, Staphylococcus aureus (n = 62), Streptococcus agalactiae (n = 46), and Escherichia coli (n = 129), were determined for dairy cattle with mastitis across 142 Ukrainian farms. The results showed that there were more gentamicin resistant S. aureus isolates (16.95%) identified in this study than previously reported for Ukrainian dairy cattle. Moreover, low levels of amoxicillin susceptibly (13.51%) were observed for St. agalactiae, which contrasted a previous study showing susceptibility levels of >50%. St. agalactiae resistance to tetracycline was observed in 80% of the isolates. Cephalosporin use was most ineffective against E. coli, with 43.27-56% of the isolates exhibiting this resistant trait. Overall, this study performed a preliminary analysis of antimicrobial resistance on mastitis isolates from Ukrainian farms. However, given the limited numbers of the isolates tested in this study and that the publications on antimicrobial resistance in animal husbandry of Ukraine are very few, more extensive investigations are needed to comprehensively examine susceptibility patterns of mastitis-causing pathogens in dairy cattle in Ukraine.

The Use of Alfaxalone in Quaker Parrots (Myiopsitta monachus).

Alfaxalone is a neurosteroid anesthetic that acts on gamma-aminobutyric acid alpha-receptors. The objective of this study was to evaluate the clinical safety and efficacy of alfaxalone (Alfaxan CD). Due to observed hyperexcitability in the subject animals when alfaxalone was the only drug used during the initial trials, premedication with midazolam was also evaluated during the final study. Ten adult Quaker parrots (Myiopsitta monachus) were assigned to 3 groups: 1) low-dose alfaxalone 10 mg/kg (LD), 2) high-dose alfaxalone 25 mg/kg (HD), and 3) alfaxalone 10 mg/ kg with midazolam 1 mg/kg premedication (AM), administered intramuscularly. Induction time, sedation quality, duration of action, and vital parameters, including heart rate, respiratory rate, and temperature, were recorded. All protocols achieved adequate sedation; however, muscle tremors and hyperexcitation were variable. The LD group had a significantly longer mean ± SD induction time (13.5 ± 4.5 minutes) as compared to the HD (6.0 ± 1.3 minutes, P = .002) and AM (6.5 ± 2.9 minutes, P = .006) groups, while recovery time was significantly longer in the HD group (86.2 ± 13.4 minutes) than the LD group (44.4 ± 10.8 minutes, P < .001). Midazolam premedication resulted in reduction of both muscle tremors and hyperexcitation associated with alfaxalone administration, but the recovery time was significantly longer (103.5 ± 15.1 minutes, P < .001) than for the LD group. Alfaxalone as a sole agent resulted in muscle tremors and hyperexcitation during induction, which was attenuated by premedication with midazolam. Further investigation is warranted to characterize the effects of alfaxalone and drugs used to premedicate Quaker parrots.

Venous Blood Gas, Electrolyte, and Hematologic Analytes of the Mottled Duck, Anas fulvigula.

We collected venous blood samples from 83 apparently healthy Mottled Ducks ( Anas fulvigula ) July 2012-August 2013 on the Texas, US, Gulf Coast and measured blood gas, electrolyte, biochemical, and hematologic parameters. Age, sex, body condition score, capture year, capture type, and time of day had significant statistical, but not clinically relevant, effects on several analytes. Ducks caught by rocket net had findings consistent with greater stress compared with hand-caught ducks. These analyte data for healthy free-living Mottled Ducks may be useful in the assessment of Mottled Duck population health and in the management and treatment of individual ducks affected by environmental stressors.

Ribavirin Inhibits Parrot Bornavirus 4 Replication in Cell Culture.

Parrot bornavirus 4 is an etiological agent of proventricular dilatation disease, a fatal neurologic and gastrointestinal disease of psittacines and other birds. We tested the ability of ribavirin, an antiviral nucleoside analog with antiviral activity against a range of RNA and DNA viruses, to inhibit parrot bornavirus 4 replication in duck embryonic fibroblast cells. Two analytical methods that evaluate different products of viral replication, indirect immunocytochemistry for viral specific nucleoprotein and qRT-PCR for viral specific phosphoprotein gene mRNA, were used. Ribavirin at concentrations between 2.5 and 25 µg/mL inhibited parrot bornavirus 4 replication, decreasing viral mRNA and viral protein load, in infected duck embryonic fibroblast cells. The addition of guanosine diminished the antiviral activity of ribavirin suggesting that one possible mechanism of action against parrot bornavirus 4 may likely be through inosine monophosphate dehydrogenase inhibition. This study demonstrates parrot bornavirus 4 susceptibility to ribavirin in cell culture.

Use of serology and bacterial culture to determine prevalence of Brucella spp. In feral Swine (sus scrofa) in proximity to a beef cattle herd positive for Brucella suis and Brucella abortus.

Using serology and bacterial culture, we determined the prevalence of Brucella spp. and the antibody to Brucella spp. in a feral swine (Sus scrofa) population in proximity to a cattle herd that was culture positive for Brucella abortus and Brucella suis in north-central Texas, USA. During a prospective cross-sectional quantitative study in April 2005, we collected blood and tissue samples from 40 feral swine within a 30-km radius of the infected herd. Serum samples were tested by the Rose Bengal test, particle concentration fluorescence immunoassay, and fluorescence polarization assay. In addition, tissue samples were cultured, and the Brucella species and biovar determined. Four feral swine were Brucella positive by serology, and two were culture positive for B. suis biovar 1. Of the culture-positive swine, one was concurrently antibody and culture positive, and one was culture positive only. The presumptive source of the B. suis infection in the index cattle herd was likely the surrounding feral swine population. Because B. abortus was not cultured from the feral swine, it is unlikely that the source of the B. abortus infection in the index herd originated from the feral swine. Endemic diseases in feral swine populations can pose a disease threat to livestock and a zoonotic risk to humans.

Pharmacokinetics after intravenous administration of flunixin meglumine in budgerigars (Melopsittacus undulatus) and Patagonian conures (Cyanoliseus patagonus).

OBJECTIVE: To investigate the disposition kinetics of flunixin meglumine when administered IV to budgerigars (Melopsittacus undulatus) and Patagonian conures (Cyanoliseus patagonus). DESIGN: Prospective cohort study. ANIMALS: 8 adult Patagonian conures and 24 adult budgerigars. PROCEDURES: Injectable flunixin meglumine (50 mg/mL) was diluted to 10 and 1. 0 mg/mL and administered IV at a dose of 5.0 mg/kg (2.3 mg/lb) to Patagonian conures and budgerigars, respectively. RESULTS: In budgerigars, the elimination half-life was 0.72 hours and the mean residence time was 0.73 hours. In Patagonian conures, the elimination half-life was 0.91 hours and the mean residence time was 1.20 hours. The concentration of flunixin was below the assay's limit of quantification (0.5 µg/mL) at 3 and 6 hours in budgerigars and Patagonian conures, respectively. A single budgerigar developed adverse effects (lethargy and signs of depression) for approximately 15 minutes following drug administration. CONCLUSIONS AND CLINICAL RELEVANCE: The half-life of flunixin in Patagonian conures and budgerigars was short following IV administration; however, results of this study suggested that IV administration of injectable flunixin meglumine at 5.0 mg/kg resulted in plasma concentrations that could potentially be anti-inflammatory and analgesic in budgerigars and Patagonian conures.

A gas chromatography-mass spectrometry assay to quantify camphor extracted from goat serum.

A sensitive gas chromatography-mass spectrometry (GC-MS) method was developed and validated for quantification and pharmacokinetics of camphor, a major monoterpene of juniper plant, in goat serum. Camphor and internal standard (terpinolene) eluates from solid phase extraction (SPE) with ethyl acetate yielded well resolved peaks and were clearly identified in total and selected ion chromatograms. The elution and injection volumes were optimized for improved detection and quantification of camphor based on peak shape, signal to noise ratio, recoveries, and repeatability. The matrix calibration curve with the good linearity (R(2)=0.998) and response in the range of 0.005-10.0 µg/mL was used to determine camphor concentration in goat serum. The GC-MS method offered sufficiently low limits of detection (1 ng/mL) and quantitation (3 ng/mL) for camphor concentration in goat serum for the pharmacokinetic study. The proposed method showed good intra- and inter-day variation with relative standard deviation (RSD) of 0.2-7.7% and produced good recovery (96.0-111.6%) and reproducibility (1.6-6.1%) at all spiked levels. Using this method on serum samples obtained from two goats orally dosed with camphor confirmed that the method is suitable for camphor studies in animals.

Evaluation of homologous and heterologous protection induced by a virulent field strain of orf virus and an orf vaccine in goats.

OBJECTIVE: To evaluate cross protection provided by administration of contagious ecthyma vaccines against strains of orf virus in goats. ANIMALS: 126 Boer-Spanish crossbred goats (3 to 20 days old). PROCEDURES: 85 goats were vaccinated with a goat-derived contagious ecthyma vaccine. Of these, 41 were challenge exposed with the virus strain for the contagious ecthyma vaccine, 40 were challenge exposed with a more virulent field strain of orf virus, and 4 were lost to predation or died. Another 41 goats were vaccinated with a vaccine produced from a more virulent field strain of orf virus; of these, 18 were challenge exposed with the virus strain of the goat-derived contagious ecthyma vaccine, 18 were challenge exposed with the more virulent field strain of orf virus, and 5 were lost to predation or died. RESULTS: Vaccination with the goat-derived contagious ecthyma vaccine did not significantly reduce the number of goats with lesions or lesion severity caused by challenge exposure with the more virulent field strain of orf virus. Vaccination with the vaccine produced from the more virulent field strain of orf virus significantly reduced the number of goats with lesions attributable to challenge exposure with the virus strain of the goat-derived contagious ecthyma vaccine, but it failed to significantly reduce lesion severity. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination did not result in cross protection for the 2 strains of orf virus. This may have been attributable to antigenic differences and may be a factor in outbreaks of contagious ecthyma in vaccinated goats.

Efficacy of an anaerobic swab transport system to maintain aerobic and anaerobic microorganism viability after storage at -80 degrees C.

An Amies agar gel swab transport system was evaluated for its ability to maintain bacterial viability and relative quantity after freezing at -80°C. Nine American Type Culture Collection (ATCC) bacterial strains were used: 3 anaerobic strains (Propionibacterium acnes, Peptostreptococcus anaerobius, and Clostridium sporogenes) and 6 facultative or strict aerobic bacterial strains (Stenotrophomonas maltophilia, Escherichia coli ([ATCC 25922 and ATCC 11775], Salmonella enterica subsp. enterica serovar Typhimurium, Staphylococcus saprophyticus, and Lactobacillus casei). The bacterial species were chosen because they corresponded to bacteria identified in psittacine feces and cloacal samples. There were no significant differences between growth scores at baseline and after storage at -80°C for 40 days for any of the bacteria examined after 48 and 72 hr of incubation, with the exception of P. anaerobius. For P. anaerobius, there was a significant reduction (P < 0.001) in the growth score after storage at -80°C for 40 days from that of the baseline; however, the bacteria were still viable. The tested swab transport system may be useful when lengthy storage and transport times necessitate freezing samples prior to culture.

Development of a contagious ecthyma vaccine for goats.

OBJECTIVE: To identify a strain of contagious ecthyma virus from goats that possesses the appropriate characteristics for an effective vaccine for goats. ANIMALS: 25 goat kids used for vaccine development and 100 goat kids used for evaluation of vaccine efficacy. PROCEDURES: 5 strains of contagious ecthyma virus were tested in a vaccination-challenge study to identify the best strain to be the seed strain for a contagious ecthyma vaccine. The vaccine derived from the chosen viral stain was tested at 2 concentrations for efficacy in a vaccination-challenge study. RESULTS: 2 of 5 viral strains induced moderate to severe scabs following infection, and 3 viral strains protected the goats from wild-type virus challenge following vaccination. Viral strain 47CE was selected as the seed source for the production of a contagious ecthyma vaccine because of the larger vaccine-to-challenge scab formation ratio. Vaccine 47CE protected all goat kids (48/48) following challenge with the wild-type contagious ecthyma virus; all goat kids (32/32) in the control group had scab formation following challenge with the wild-type contagious ecthyma virus, which indicated no protection following administration of vaccine diluent. CONCLUSIONS AND CLINICAL RELEVANCE: A vaccine containing a caprine strain of contagious ecthyma virus used in goats appeared to provide the characteristics needed for an effective vaccine, including good scab production and protection from wild-type infection. This vaccine may potentially provide better protection for goats from contagious ecthyma than currently available vaccines labeled for sheep.

A practitioner's primer on foot-and-mouth disease.

Foot-and-mouth disease (FMD) is caused by an RNA virus of the genus Aphthovirus; 7 immunologically distinct serotypes of the virus have been identified. Susceptible species are mainly domestic and wild even-toed ungulates, such as cattle, sheep, goats, pigs, bison, and deer. All body fluids of infected animals can contain the virus and are considered infective. The primary mode of transmission is animal-to-animal transmission through inhalation or ingestion of aerosols containing the virus. The virus can also be spread mechanically by contaminated organic debris and fomites and can survive for 48 hours on human oral and nasal mucosa and be spread to uninfected animals in this manner. There is a rapid progression of clinical signs after an animal becomes infected, and the virus spreads rapidly throughout a herd. Clinical signs include excessive salivation; fever; vesicles and erosions of the oral and nasal mucosa, coronary band, interdigital area, and teats; lameness; sloughing of claws; reluctance to move; anorexia; mastitis; decreased milk production; and abortion or weak newborns. In mature animals, FMD has high morbidity and low mortality rates. Infected animals can become inapparent carriers of the virus.

Comparison of microbial growth inhibition screening assays with high-pressure liquid chromatography for detection of experimentally incurred penicillin G residues in calves.

Twelve calves were fed milk replacer containing 3.33 ppm penicillin G for one feeding, and 12 calves were fed the medicated milk replacer once daily at a rate of 12% of their body weight for 14 days. Two calves served as controls. Calves were slaughtered 4 to 13 hours after being fed. Kidney, liver, muscle, and urine samples were assayed for penicillin by high-pressure liquid chromatography (HPLC). Penicillin G was not detected in any muscle samples and concentrations of penicillin exceeded the tolerance level (0.05 microg/g) in kidney or liver tissue from 13 of 24 treated-calf carcasses examined by HPLC. The Live Animal Swab Test identified all 13 carcasses with violative drug residues. Using kidney as the test matrix, the Swab Test on Premises identified 10 of 13 carcasses with violative drug residues, while the Calf Antibiotic Sulfa Test identified seven of 13 carcasses with violative residues.