Successful treatment of suspected exertional myopathy in a rhea (Rhea americana).

A 7-yr-old, adult, female greater rhea (Rhea americana) from the National Zoological Park presented with a 24-hr history of severe left leg lameness that progressed to an inability to stand. Blood work revealed creatine phosphokinase (CPK) above 50,000 U/L and elevated lactate dehydrogenase. The bird's condition deteriorated over the next week. The bird's CPK increased to over 208,400 U/L. Aggressive intravenous fluids and physical therapy along with oral anxiolytic and muscle-relaxant drugs were instituted. After 2 wk of aggressive therapy, initial signs of improvement were noted. By day 28, the bird was able to walk unassisted with no noticeable lameness. This is one of the few reported cases of successful treatment of suspected ratite exertional myopathy. It is believed that success in this case can be attributed to persistent, aggressive physical therapy, muscle relaxants, and anxiolytics aimed to counteract the hyperexcitable nature of these birds.     

Development of an endogenous virus-free line of chickens susceptible to all subgroups of avian leukosis virus

Primary chicken embryo fibroblasts (CEF) from special specific pathogen-free chicken lines are used for detection of contamination of adult or embryonic tissues, meconium, or tissue culture fluids with avian leukosis viruses (ALV). The suitability and efficiency of such tests depend on the susceptibility of CEF to the various subgroups of exogenous as well as endogenous ALV. The ideal CEF for such tests should be not only susceptible to all retroviruses, but also free of endogenous viruses so that such tests are immune to any interference that may occur between the endogenous and the tested (exogenous) viruses. CEF and/or chickens free of endogenous viruses are also desirable for gene transfer studies using retroviral vectors, such as RNA interference (RNAi) experiments and transgenic work. The absence of ev genes in CEF or chickens can empower clean detection of successful RNAi construct delivery or gene transfer. CEF free of ev genes are also essential reagents routinely used in growing and detecting unknown retroviruses in varied viral assays. This report documents the development of a new line of chickens, 0.TVB*S1, that is free of endogenous viruses and susceptible to all subgroups of ALV identified in chickens.     

Comparison of the measurement of total carbon dioxide and strong ion difference for the evaluation of metabolic acidosis in diarrhoeic calves

Eighty-four calves with diarrhoea were treated with fluids and 13 apparently healthy calves of similar ages were sampled as controls. Their total blood carbon dioxide (TCO2) was measured with a Harleco apparatus and 31 of the calves were treated with oral fluids and 53 with parenteral fluids. The oral fluid contained 118 mmol/litre Na+, 25 mmol/litre K+, 110 mmol/litre glucose, 108 mmol/litre bicarbonate (HCO3- as citrate), 43 mmol/litre Cl-, 4 mmol/litre Ca++, 4 mmol/litre Mg++ and 20 mmol/litre glycine, and the parenteral fluid contained 144 mmol/litre Na+, 4 mmol/litre K+, 35 mmol/litre HCO3- and 113 mmol/litre Cl-. Both treatments resulted in significant improvements in acid-base status as demonstrated by an increase in TCO2, and the treatment was successful in 27 of the 31 calves receiving oral fluids and in 45 of the 53 calves receiving parenteral fluids. Thirty-seven of the calves treated parenterally were very severely acidotic (TCO2 <8 mmol/litre) initially and they received an additional 400 mmol HCO3- added to the first 5 litres of infusion. Treatment was successful in 33 of these calves. The decision to administer additional bicarbonate was made on the basis of their acid-base status as measured with a Harleco apparatus. The strong ion difference (Na++K+-Cl-) (SID) of the calves was calculated retrospectively. There was a significant correlation between the SID and TCO2 of the calves treated with oral fluids but not among the control calves or the calves treated parenterally. Furthermore, measurements of the change in SID during therapy gave little indication of the change in acid-base status as measured by the Harleco apparatus, with the SID decreasing (suggesting a worsening of acid-base status) in 16 calves in which the TCO2 increased (suggesting an improvement in acid-base status). There was a significant correlation between the change in SID and the change in TCO2 during treatment in the calves receiving oral fluids but not in the calves treated parenterally.     

Oral Recombinant Feline Interferon-Omega as an alternative immune modulation therapy in FIV positive cats: Clinical and laboratory evaluation

Recombinant-Feline Interferon-Omega (rFeIFN-ω) is an immune-modulator licensed for use subcutaneously in Feline Immunodeficiency virus (FIV) therapy. Despite oral protocols have been suggested, little is known about such use in FIV-infected cats. This study aimed to evaluate the clinical improvement, laboratory findings, concurrent viral excretion and acute phase proteins (APPs) in naturally FIV-infected cats under oral rFeIFN-ω therapy (0.1 MU/cat rFeIFN-ω PO, SID, 90 days). 11 FIV-positive cats were treated with oral rFeIFN-ω (PO Group). Results were compared to previous data from 7 FIV-positive cats treated with the subcutaneous licensed protocol (SC Group). Initial clinical scores were similar in both groups. Independently of the protocol, rFeIFN-ω induced a significant clinical improvement of treated cats. Concurrent viral excretion and APP’s variation were not significant in the PO Group. Oral rFeIFN-ω can be an effective alternative therapy for FIV-infected cats, being also an option for treatment follow-up in cats submitted to the licensed protocol.     

Clindamycin bioavailability and pharmacokinetics following oral administration of clindamycin hydrochloride capsules in dogs

Oral bioavailability and pharmacokinetic behaviour of clindamycin in dogs was investigated following intravenous (IV) and oral (capsules) administration of clindamycin hydrochloride, at the dose of 11 mg/kg BW. The absorption after oral administration was fast, with a mean absorption time (MAT) of 0.87+/-0.40 h, and bioavailability was 72.55+/-9.86%. Total clearance (CL) of clindamycin was low, after both IV and oral administration (0.503+/-0.095 vs. 0.458+/-0.087 L/h/kg). Volume of distribution at steady-state (IV) was 2.48+/-0.48 L/kg, indicating a wide distribution of clindamycin in body fluids and tissues. Elimination half-lives were similar for both routes of administration (4.37+/-1.20 h for IV, vs. 4.37+/-0.73 h for oral). Serum clindamycin concentrations following administration of capsules remained above the MICs of very susceptible microorganisms (0.04-0.5 microg/mL) for 12 or 10 h, respectively. Time above the mean inhibitory concentration (MIC) is considered as the index predicting the efficacy of clindamycin (T(>MIC) must be at least 40-50% of the dosing interval), so a once-daily oral administration of 11 mg/kg BW of clindamycin can be considered therapeutically effective. For less susceptible bacteria (with MICs of 0.5-2 microg/mL) the same dose should be given but twice daily.     

Synthesis and detection of toltrazuril sulfone and its pharmacokinetics in horses following administration in dimethylsulfoxide

Triazine-based antiprotozoal agents are known for their lipophylic characteristics and may therefore be expected to be well absorbed following oral administration. However, although an increase in lipid solubility generally increases the absorption of chemicals, extremely lipid-soluble chemicals may dissolve poorly in gastrointestinal (GI) fluids, and their corresponding absorption and bioavailability would be low. Also, if the compound is administered in solid form and is relatively insoluble in GI fluids, it is likely to have limited contact with the GI mucosa, and therefore, its rate of absorption will be low. Based on the above considerations, we sought a solvent with low or no toxicity that would maintain triazine agents in solution. As the oral route is most preferred for daily drug therapy, such a solvent would allow an increased rate of absorption following oral administration. In present study, it was demonstrated that dimethylsulfoxide (DMSO) increased the oral bioavailability of toltrazuril sulfone (Ponazuril) threefold, relative to oral administrations of toltrazuril sulfone suspended in water. The cross-over study of toltrazuril sulfone formulated in DMSO indicated that the absolute oral bioavailability of toltrazuril sulfone in DMSO is 71%. The high bioavailability of the DMSO-preparation suggests that its daily oral administration will routinely yield effective plasma and cerebral spinal fluid (CSF) concentrations in all horses treated. Also, this improved formulation would allow clinicians to administer loading doses of toltrazuril sulfone in acute cases of Equine Protozoal Myeloencephalitis. Another option would involve administration of toltrazuril sulfone in DMSO mixed with feed (1.23 kg daily dose) meeting the US Food and Drug Administration (FDA) recommendations for the levels of DMSO permissible in pharmaceutical preparations.     

Oral fluid samples used for PRRSV acclimatization program and sow performance monitoring in endemic PRRS-positive farms

An effective gilt acclimatization program is one of the most important management strategies for controlling porcine reproductive and respiratory syndrome virus (PRRSV) infection. Recently, oral fluid samples have been used as alternative diagnostic samples for various swine diseases. This study utilized oral fluids for PRRSV monitoring during the gilt acclimatization period in PRRSV endemic farms. The study was performed in two selected commercial breeding herds (farm A and farm B). PRRSV RNA and PRRSV-specific antibodies were monitored using oral fluid and serum samples. Sow performance parameters related to PRRSV infection were recorded and assessed. After PRRSV exposure during acclimatization, viral RNA was demonstrated in oral fluids from 1 to 10 weeks post-exposure (WPE). PRRSV RNA was detected in serum at 1 and 4 WPE in farm A and at 1, 4, 8, and 12 WPE in farm B. Prolonged viremia of gilts from farm B was possibly due to re-infection (within the herd) and later, reproductive problems were found in the breeding herd. The correlation of PRRSV RNA concentration in oral fluids and serum was evident. The S/P ratio values of PRRSV antibodies in oral fluid samples were higher and had similar patterns of antibody responses to the serum samples. The results suggest that the use of oral fluid samples for PRRSV monitoring during gilt acclimatization in endemic farms is effective, convenient, practical, and economical and would be most beneficial when used with other parameters.     

Presence of Oxytocin,Vasopressin and Atrial Natriuretic Peptide and Their Modification in Rat Hypothalamic Paraventricular Nucleus During Resistance Training

Many studies have demonstrated the physiological effects of oxytocin (OT), atrial natriuretic peptide (ANP) and vasopressin (VP) in the homoeostasis of body fluids during physical exercise. However, a little information is available about the related immunohistochemical changes in hypothalamic magnocellular neurosecretory system during and after the training. The aim of the present work was to study the immunohistochemical changes in OT, ANP and VP levels in the hypothalamic paraventricular nucleus during and after resistance exercise protocol. Three groups of Wistar rats were trained by a rung ladder protocol for 15, 30 and 45 days, respectively; a fourth group was left to rest for 15 days after the training. Finally, four sedentary groups were used as controls. The results show that resistance training induces a significant reduction in the percentage of OT‐positive neurons, compared with sedentary controls. In contrast, this protocol did not induce any change in VP levels, and ANP levels did not change significantly. However, VP increased after the resting period of 15 days. Our work shows that neurons of the paraventricular nucleus are involved in body fluid homoeostasis during and after resistance exercise. The functional significance of these changes in OT and VP levels, during and after the protocol, needs to be further investigated.     

Porcine reproductive and respiratory syndrome virus whole-genome sequencing efficacy with field clinical samples using a poly(A)-tail viral genome purification method

The genomic surveillance of porcine reproductive and respiratory syndrome virus (PRRSV) is based on sequencing of the ORF5 gene of the virus, which covers only 4% of the entire viral genome. It is expected that PRRSV whole-genome sequencing (WGS) will improve PRRSV genomic data and allow better understanding of clinical discrepancies observed in the field when using ORF5 sequencing. Our main objective was to implement an efficient method for WGS of PRRSV from clinical samples. The viral genome was purified using a poly(A)-tail viral genome purification method and sequenced using Illumina technology. We tested 149 PRRSV-positive samples: 80 sera, 33 lungs, 33 pools of tissues, 2 oral fluids, and 1 processing fluid (i.e., castration liquid). Overall, WGS of 67.1% of PRRSV-positive cases was successful. The viral load, in particular for tissues, had a major impact on the PRRSV WGS success rate. Serum was the most efficient type of sample to conduct PRRSV WGS poly(A)-tail assays, with a success rate of 76.3%, and this result can be explained by improved sequencing reads dispersion matching throughout the entire viral genome. WGS was unsuccessful for all pools of tissue and lung samples with Cq values > 26.5, whereas it could still be successful with sera at Cq ≤ 34.1. Evaluation of results of highly qualified personnel confirmed that laboratory skills could affect PRRSV WGS efficiency. Oral fluid samples seem very promising and merit further investigation because, with only 2 samples of low viral load (Cq = 28.8, 32.8), PRRSV WGS was successful.     

An efficient multiplex PCR suitable for large scale typing in linkage mapping.

The dissection of polygenic traits is made possible with the development of microsatellite markers. Linkage study of this kind involves many markers with tens of hundreds of samples. Although typing essentially contains only two steps: PCR amplification and gel electrophoresis. Such work is still heavy when a large number of samples had to be genotyped. Multiplex PCR may reduce the work, but one has to optimize the conditions from marker to marker. Here we describe a dye-compatible multiplex PCR that works under standardized condition without the need to pre-determine the combinational primer concentration and the time-consuming step to mix many samples with gel loading dye before electrophoresis. This successful protocol should greatly reduce the cost and labor for genetic study of polygenic traits.     

An appropriate ingestion volume of oral sulfa drug suspension in pigs

The influence of ingested volume of a sulfa drug suspension, sodium sulfamonomethoxine (SMMNa), on the oral pharmacokinetics was studied in pigs, with regard to bioavailability and gastric emptying. Eighteen pigs, weighing 30-70 kg, were used. Phenol red solution was used for the evaluation of gastric emptying study. SMMNa suspension was used for pharmacokinetic study. Both of these fluids were administered by natural swallowing. Three experimental groups were constructed: G-I; 5 ml/kg of the test fluids to starved animals, G-II; 5 ml/kg of the test fluids to fed animals and G-III; 20 ml/kg of the fluids to fed animals. The glucose glycine electrolyte solution (GGES) was used as the vehicle for both the compounds. Six pigs, having duodenal cannula, were used for the study of gastric emptying. The gastric emptying rate was rapid in G-I, relatively rapid in G-III, and slow and variable in G-II. In agreement with the result of gastric emptying study, the values of Cmax and tmax were high and rapid in G-I, relatively high and rapid in G-III, and low and slow in G-II. Accordingly, the voluminous ingestion of drug suspension can facilitate the gastric emptying, in turn may make the oral absorption of the drug rapid-and-uniform. The 20 ml/kg volume of sulfa drug suspension may practically be recommended for the oral administration in pigs.     

Eosinophils of the horse: Part II: Eosinophils in clinical diseases

Interpretation of eosinophilia in body fluids or tissues is often not straightforward. Eosinophil counts vary among clinically healthy individuals, and considerable overlap can occur between normal and affected animals in conditions such as allergic airway disease. Parasite exposure is a confounding factor when counts are increased, and in cases where very high counts and dramatic clinical signs make another disease process obvious, the underlying pathology may be uncertain and treatment difficult. Eosinophils are a component of the immune response in many diseases of the horse, but their specific role is often unknown and likely multifactorial. In helminth infections, eosinophils are assumed to be part of the normal host response to a pathogen, whereas in multisystemic eosinophilic epitheliotropic disease (MEED), the predominance of eosinophils likely represents a wildly dysregulated response, or an abnormal response altogether. This distinction is still not clear for other diseases. Understanding the pathways involved in recruitment, activation or suppression of eosinophils is required for more accurate diagnostics, effective therapeutics, and successful strategies for prevention of eosinophil associated diseases. Eosinophils of the horse: Part II reviews published observations on the eosinophil in clinical diseases of the horse. The behaviour of eosinophils in three common and relatively well-studied conditions is presented first, including gastrointestinal helminth infections, non-infectious respiratory disease, and insect bite hypersensitivity. The less common eosinophil-associated diseases such as eosinophilic disease confined to the intestine (EDCI) and MEED are considered, followed by a brief summary of the eosinophil in phycomycosis and neoplasia. In conclusion, a panoramic view of the equine eosinophil as presented in Parts I and II is placed in the larger context of current eosinophil research, and areas of study are identified that may improve our understanding of eosinophil biology in equine health and clinical disease.     

Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral,tonsillar, and nasal fluids

Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control.     

Owner Compliance to an Environmental Management Protocol for Severe Equine Asthma Syndrome

Severe equine asthma (SEA) syndrome is a chronic recurrent respiratory disease, common among adult horses. The disease occurs in genetically susceptible individuals after their exposure to organic dust. Thus, environmental management has proved essential in controlling airway challenge and disease exacerbation. This is a demanding process that can only be achieved through the horse owners’ cooperation. One year after initial diagnosis of SEA in a group of 39 horses, owner compliance to an environmental management protocol was evaluated. The overall compliance to the protocol was poor and the horses’ clinical health and need for pharmacological management was related to the successful implementation of the environmental recommendations provided on disease diagnosis.     

An optimized high quality male DNA extraction from spermatophores in open thelycum shrimp species

The crucial step of most of the current genetic studies is the extraction of DNA of sufficient quantity and quality. Several genomic DNA isolation methods have been described to successfully obtain male DNA from shrimp species. However, all current protocols require invasive handling methods with males for DNA isolation. Using Aristeus antennatus as a model we tested a reliable non‐invasive differential DNA extraction method to male DNA isolation from spermatophores attached to female thelycum. The present protocol provides high quality and quantity DNA for polymerase chain reaction amplification and male genotyping. This new approach could be useful to experimental shrimp culture to select sires with relevant genetic patterns for selective breeding programs. More importantly, it can be applied to identify the mating pairs and male structure in wild populations of species as A. antennatus, where males are often difficult to capture. Our method could be also valuable for biological studies on other spermatophore‐using species, such as myriapods, arachnids and insects.     

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.     

Antibodies to immunoglobulin-G in dog sera, synovial fluids and aqueous humor: a comparative study of rheumatoid factor assays, suitable for routine application

The incidence of anti-IgG antibodies (rheumatoid factors, RF) in body fluids (sera, synovial fluids and aqueous humor) selected from 62 normal and 275 diseased dogs was studied. Fluids were assayed by canine versions of standard agglutinating and/or precipitating RF assays with routine application in human practice. The number of RF detected by dog IgG-coated particles was substantially higher by latex fixation test (LFT) than by modified Rose-Waaler (RW) test (61/144 vs. 14/144). This did not result from false positives by LFT since latex activity was completely inhibited by aggregated dog IgG. Some evidence is presented indicating that results obtained by standard RW in particular, but also those obtained by standard LFT, might be improved by modifying testing conditions currently used. Body fluids were further studied for the presence of precipitins to aggregated dog IgG in 0.6% agarose (gel precipitation test (GPT]. The frequency of RF was higher by GPT than by LFT, both in normal control fluids (for sera 26/52 vs. 19/52) and patient material (for sera 135/197 vs. 95/197). Thus, the canine RF appear to be a serum component with an unexpectedly high frequency in both normal and diseased dogs, but grossly underestimated by the recommended routine RF assays based on agglutination. The GPT, which combines a superior detection rate of theoretically also agglutinating RF with an inability to detect RF quantitatively, seems an ideal RF 'indicator' test to dictate improvements to the quantitative LFT/RW assays so as to facilitate RF detection at clinically relevant concentrations. Thus optimized, RW/LFT would provide the optimal detection apparatus for the ultimate isolation of the relevant 'RF' repertoire present, for comparative studies aimed ultimately at unraveling the etiopathogenesis of the 'real' RF.     

Oral administration of lactoferrin inhibits inflammation and nociception in rat adjuvant-induced arthritis

Lactoferrin (LF) is a ubiquitous protein which exists in milk, plasma, synovial fluids, cerebrospinal fluid and other biological fluids. LF is also well known as a natural immunomodulator. Recently, we found that bovine milk-derived LF (BLF) produced micro-opioid receptor-mediated analgesia. In this study, we examined whether oral administration of BLF causes anti-nociceptive and anti-inflammatory effects, and also whether it modulates LPS-induced TNF-alpha and IL-10 production in rat model of rheumatoid arthritis (RA), rat adjuvant arthritis. BLF was administrated once daily, starting 3 hr before (preventive experiment) or 19 days after (therapeutic experiment) adjuvant injection. In both experiments, BLF suppressed the development of arthritis and the hyperalgesia in the adjuvant-injected paw. The single-administered BLF produced a dose-dependent analgesia, which was reversed by naloxone, in the adjuvant arthritis rats. Both repeated and single administration of BLF suppressed TNF-alpha production and increased IL-10 production in the LPS-stimulated adjuvant arthritis rats. These results suggest that orally administered BLF has both preventive and therapeutic effects on the development of adjuvant-induced inflammation and pain. Moreover, the immunomodulatory properties of BLF, such as down-regulation of TNF-alpha and up-regulation of IL-10, could be beneficial in the treatment of RA. Thus, we concluded that LF can be safely used as a natural drug for RA patients suffering from joint pain.     

Efficacy of oseltamivir phosphate to horses inoculated with equine influenza A virus

We investigated the efficacy of the oral administration of oseltamivir phosphate (OP) in horses experimentally infected with equine influenza A virus (H3N8). Nine horses were divided into three horses each of control, treatment and prophylaxis groups. An administration protocol for the treatment group (2 mg/kg of body weight, twice a day for five days) was started immediately after the onset of pyrexia (above 38.9 degrees C). An administration protocol for the prophylaxis group (2 mg/kg of body weight, once a day for five days) was started on a day before viral inoculation. In the treatment group, periods of virus excretion (mean days +/- standard deviation, 2.3 +/- 0.6) and pyrexia (2.0 +/- 0.0) were apparently shorter than those of the control group (6.0 +/- 0.0 and 8.0 +/- 1.0, respectively). In the prophylaxis group, although virus excretion and pyrexia were not prevented, the periods of virus excretion (5.0 +/- 0.0) and pyrexia (4.7 +/- 1.5) were shorter than those of the control group. Moreover, in the treatment and prophylaxis groups, bacterial counts of Streptococcus equi subsp. zooepidemicus known as the common pathogen of secondary bacterial pneumonia in bronchoalveolar lavage fluids collected seven days after inoculation were significantly fewer than that of the control group. The results indicated that the oral administration of OP to horses affected with equine influenza would contribute to reduce the magnitude of virus excretion, pyrexia and consequent secondary bacterial pneumonia.