Expression of inducible nitric oxide synthase in macrophages inversely correlates with parasitism of lymphoid tissues in dogs with visceral leishmaniasis

Background

There are only a few studies reporting the role of nitric oxide metabolites for controlling macrophage intracellular parasitism, and these are controversial. Therefore, the present study aimed to evaluate the expression of inducible nitric oxide synthase (iNOS) in the lymph nodes and spleen of dogs affected by visceral leishmaniasis through immunohistochemistry and to determine its correlation with tissue parasite burden and serum interferon (IFN)-γ levels. Twenty-eight dogs were selected and assigned to one of two groups, symptomatic (n = 18) and asymptomatic (n = 10), according to clinical status and laboratory evaluation. A negative control group (n = 6) from a non-endemic region for visceral leishmaniasis was included as well.

Results

Parasite density (amastigotes/mm²) was similar between clinical groups in the lymph nodes (P = 0.2401) and spleen (P = 0.8869). The density of iNOS⁺ cells was higher in infected dogs compared to controls (P < 0.05), without a significant difference in lymph node (P = 0.3257) and spleen (P = 0.5940) densities between symptomatic and asymptomatic dogs. A positive correlation was found between the number of iNOS⁺ cells in lymph nodes and interferon-γ levels (r = 0.3776; P = 0.0303), and there was a negative correlation between parasites and iNOS⁺ cell densities both in lymph nodes (r = −0.5341; P = 0.0034) and spleen (r = −0.4669; P = 0.0329).

Conclusion

The negative correlation observed between tissue parasitism and the expression of iNOS may be a reflection of NO acting on the control of parasites.     

Antimicrobial delivery by intrasynovial catheterisation with systemic administration for equine synovial trauma and sepsis

Objective To present the technique for intra-articular catheter placement and report the clinical outcomes of 38 cases of equine synovial trauma and/or infection treated with broad-spectrum antimicrobials administered via an intrasynovial catheter (ISC). Design Retrospective study. Procedure Medical records of 38 horses treated for synovial trauma and sepsis with frequent antimicrobial administration through an ISC from 1995 to 2008 were reviewed. Follow-up information was obtained via clinical re-evaluation or telephone contact with the owners. Results The majority of horses (84%) received amikacin and Timentin® four times daily. In addition, synovial lavage through the ISC was carried out in 27 horses (71%). Only radiological evidence of osteolysis had a significant negative impact on both lameness at the time of hospital discharge and the long-term outcome. In total, 92% of horses treated with frequent antimicrobial administration through an ISC had clinical resolution of infection. Catheter obstruction occurred in three cases, necessitating replacement or removal, and two synovial fistulae developed at sites of open drainage. The majority of horses treated had a favourable outcome, with 86% being at least pasture sound and 43% returned to riding. Conclusion Septic synovial structures treated with frequent antimicrobial administration through an ISC had a good prognosis for survival and 43% returned to riding, which is consistent with the results of other studies. The use of a simple ISC should be considered when broad-spectrum intrasynovial antimicrobial administration and lavage of a septic synovial structure are indicated.     

Meningoencephalomyelitis in a foal due to Salmonella agona infection

CASE HISTORY: A neonatal Thoroughbred foal was presented with rib fractures and left forelimb lameness secondary to dystocia.

CLINICAL FINDINGS: The foal developed a head tilt, seizures and watery diarrhoea during hospitalisation and died at 7 days of age. Histological examination of the brain and spinal cord revealed a suppurative meningoencephalomyelitis with vasculitis, and numerous intralesional, gram-negative bacilli. Similar microscopic lesions were noted in the lungs, renal medullary interstitium, and umbilicus. Bacilli in the brain, spinal cord and umbilicus were identified immunohistochemically as Salmonella group B. Salmonella agona was isolated in pure culture from the brain, lung, liver, kidney, and intestine.

CONCLUSION: This is the first report of meningoencephalomyelitis and septicaemia due to Salmonella infection in an equine neonate.     

Systemic Immunosuppression by Methylprednisolone and Pregnancy Rates in Goats Undergoing the Transfer of Cloned Embryos

The presence of the zona pellucida has been perceived as a requirement for the oviductal transfer of cloned embryos at early stages of development while protecting the embryo from an immune system response. We hypothesized that steroid hormone therapy could reduce a potential cellular immune response after the transfer of zona‐free cloned embryos into the oviduct of recipient female goats. In Experiment 1, seven does were used to study the systemic immunosuppressant effect of the methylprednisolone administration (for 3 days) on blood cell counts. Whole blood was collected prior to treatment with methyprednisolone and then on Days 1, 2, 3, 4, 7, 14, 21 and 28 after the first dose of methylprednisolone for the analysis of haematological parameters. Methylprednisolone treatment significantly reduced circulating white blood cells and neutrophils in comparison with pre‐treatment levels, demonstrating a systemic immunosuppressant effect. In Experiment 2, a group of 58 does were used as recipient females to study the effect of administration of methylprednisolone for 3 days on the establishment of pregnancies after the transfer of zona‐free cloned embryos into the oviducts. No effects on pregnancy rates on Day 30 were observed regarding the distinct treatment groups (control vs. methylprednisolone), the source of oocytes (in vivo‐ vs in vitro‐matured) or the presence or absence of the zona pellucida in embryos. In summary, methylprednisolone was effective at inducing a systemic immunosuppressed state in goats, but the treatment prior to embryo transfer did not affect pregnancy rates. Moreover, pregnancy rates were similar between zona‐free and zona‐intact goat cloned embryos.     

Effect of body condition score and reuse of progesterone‐releasing intravaginal devices on conception rate following timed artificial insemination in Nelore cows

This study had the aim of investigating the efficiency of timed artificial insemination (TAI) through the progesterone‐releasing intravaginal device (PRID), used in new condition and for the second and third times in Nelore cows. The effects of device reuse and body condition score (BCS) on the conception rate (CR) were evaluated in 1,122 multiparous Nelore cows (mean BCS of 2.7 ± 0.4), which were randomly distributed into three groups that received new (n = 330), once (n = 439) and twice used (n = 353) PRID. Among the 1,122 females that underwent TAI, 573 became pregnant, thus representing an overall CR of 51.06%. Cows with BCS between 2.75 and 4.0 had greater (p < .0001) CR (69.75%) than cows with BSC of 2.0–2.5 (32.98%). It was observed that the CR through using PRID was 60.00%, 51.71% and 41.93% for new, once and twice used PRID, respectively, with difference between all groups (p < .0001). Under tropical conditions, animals with BCS greater than 2.5 had a higher CR, and the CR decreased proportionally with the number of times that the PRID had been used.     

Short‐Term Antiandrogen Flutamide Treatment Causes Structural Alterations in Somatic Cells Associated with Premature Detachment of Spermatids in the Testis of Pubertal and Adult Guinea Pigs

In spite of widespread application of flutamide in the endocrine therapies of young and adult patients, the side effects of this antiandrogen on spermatogenesis and germ‐cell morphology remain unclear. This study evaluates the short‐term androgen blockage effect induced by the administration of flutamide to the testes of pubertal (30‐day old) and adult (65‐ and 135‐day old) guinea pigs, with an emphasis on ultrastructural alterations of main cell types. The testes removed after 10 days of treatment with either a non‐steroidal antiandrogen, flutamide (10 mg/kg of body weight) or a pharmacological vehicle alone were processed for histological, quantitative and ultrastructural analysis. In pubertal animals, flutamide androgenic blockage induces spermatogonial differentiation and accelerates testes maturation, causing degeneration and detachment of primary spermatocytes and round spermatids, which are subsequently found in great quantities in the epididymis caput. In post‐pubertal and adult guinea pigs, in addition to causing germ‐cell degeneration, especially in primary spermatocytes, and leading to the premature detachment of spherical spermatids, the antiandrogen treatment increased the relative volume of Leydig cells. In addition, ultrastructural evaluation indicated that irrespective of age antiandrogen treatment causes an increase in frequency of organelles involved with steroid hormone synthesis in the Leydig cells and a dramatic accumulation of myelin figures in their cytoplasm and, to a larger degree, in Sertoli cells. In conclusion, the transient exposition of the guinea pigs to flutamide, at all postnatal ages causes some degenerative lesions including severe premature detachment of spermatids and accumulation of myelin bodies in Leydig and Sertoli cells, compromising, at least temporarily, the spermatogenesis.     

Increased yearling weight as a proportion of 21-month weight was associated with increased milk production in dairy heifers

ABSTRACT

Aims: To examine the relationship between liveweight (LWT) at 12 months as a proportion of LWT at 21 months of age (LWT(12/21)%) and first lactation and cumulative 3-year milk production in dairy heifers in New Zealand.

Methods: Liveweight and milk production records were obtained for dairy heifers born from June to December (spring-calving season) between 2006–2007 and 2013–2014 dairy seasons; production records included first lactation (n?=?140,113) and cumulative 3-year (n?=?67,833) milksolids and energy-corrected milk (ECM) yields. Heifers were classified into five breed groups; Holstein-Friesian, Holstein-Friesian crossbred, Jersey, Jersey crossbred and Holstein-Friesian-Jersey crossbred. Within each breed group heifers were categorised into quintiles based on 21-month LWT. The LWT(12/21)% was calculated for each animal. Relationships between LWT(12/21)% and milk production within each breed group and LWT category were estimated using linear mixed effects models including the linear and quadratic effects of LWT(12/21)%.

Results: The relationship between LWT(12/21)% and milk production was predominantly curvilinear, with lower milk production at lesser LWT(12/21)% compared with greater LWT(12/21)%. For all breed groups and most LWT categories, heifers that were 55 or 65% LWT(12/21)% produced greater ECM and milksolids yields compared with heifers that were 45% LWT(12/21)%. Holstein-Friesian, Holstein-Friesian crossbred and Holstein-Friesian-Jersey crossbred heifers that were 65% LWT(12/21)% produced greater cumulative 3-year ECM and milksolids yields compared with heifers of the same breed group that were 45% LWT(12/21)%

Conclusions and clinical relevance: Heifers that were a greater proportion of their 21-month LWT at 12 months of age produced more first lactation and cumulative 3-year milk yields than heifers that were a lesser proportion of their 21-month LWT at 12 months of age. These results indicate that increased growth in early life of New Zealand dairy heifers is beneficial to future milk production.     

The evaluation of endophyte toxin residues in sheep fat

Abstract

AIM: To monitor changes in concentrations of lolitrem B and epoxy-janthitrems in the fat of sheep grazing perennial ryegrass infected with wild-type- and AR37-endophyte, respectively, during the time of year when ryegrass staggers would be expected to be observed.

METHODS: Ten 5-month-old lambs with no previous exposure to endophytes were grazed on either wild-type (containing lolitrem B, n = 5) or AR37 (containing epoxy-janthitrems, n = 5) endophyte-infected perennial ryegrass (Lolium perenne L.) pastures between October 2008 and June 2009. Animals were regularly assessed for ryegrass staggers using the Keogh scale (0 = no signs, 5 = severe tremors). When a score of > 3.5 was observed animals were removed from the treatment pastures for 1 month. Fat biopsy samples were taken from each animal at approximately monthly intervals and analysed for endophyte metabolites using high performance liquid chromatography (HPLC) methods developed during this study. Regular herbage samples were also taken and concentrations of endophyte metabolites measured.

RESULTS: Efficient and reproducible methods to analyse both lolitrem B and epoxy-janthitrems in fat were developed. Concentrations of lolitrem B and epoxy-janthitrems in herbage and in sheep fat increased from late November to peak in mid-February. Ryegrass staggers was observed in both groups of sheep at this time. Following 1 month of grazing non-infected pasture mean concentrations in fat of lolitrem B decreased by 43% from 61.8 to 35.3 ppb, and of epoxy-janthitrems by 38% from 1032.0 to 639.5 ppb. Maximum concentrations in herbage of epoxy-janthitrems (35.7 ppm) were higher than of lolitrem B (3.4 ppm), but signs of staggers were less severe in sheep grazing pasture containing the former compared with the latter (median Keogh scores in late February were 2 and 3, respectively), consistent with epoxy-janthitrems being low potency toxins.

CONCLUSION: This study demonstrated that concentrations of epoxy-janthitrems and lolitrem B in sheep fat increased quickly during the initial phase of the study when concentrations in pasture increased, and decreased when animals were removed from pastures containing these compounds. These data will be used in the risk assessment of the endophyte metabolites.