Genetic characterisation of Giardia duodenalis in dairy cattle in Brazil

The intestinal protozoan parasite Giardia duodenalis (Lambl, 1859) Kofoid & Christiansen, 1915 [syn. Giardia intestinalis and Giardia lamblia] has emerged as a widespread enteric pathogen in humans and domestic animals. In recent years, G. duodenalis has been found in cattle worldwide and longitudinal studies have reported cumulative prevalence of 100% in some herds. In the present study, we determined the prevalence and genetic characterisation of G. duodenalis in 200 dairy cattle from 10 dairy farms in S?o Paulo state, Brazil. All faecal specimens were screened for the presence of G. duodenalis using microscopy examination, enzyme immunoassay (EIA) and polymerase chain reaction (PCR). DNA was extracted from faecal samples and G. duodenalis were identified by amplification of the small subunit ribosomal (SSU-rDNA) and glutamate dehydrogenase (GDH) genes followed by restriction fragment length polymorphism (RFLP) or sequencing analysis. Giardia was identified in eight farm locations (80% prevalence). Overall, 15/200 (7.5%) animals were positive for infection, only one of which was a cow. Giardia duodenalis genotype E was present in 14 of the animals tested. Zoonotic genotype AI was present in one positive sample. Genotype E and genotype A represented 93% and 7% of G. duodenalis infections, respectively. This study demonstrates that G. duodenalis infection was prevalent in dairy calves in S?o Paulo state and that the non-zoonotic genotype E predominates in cattle in this region. Nevertheless, calves naturally infected in Brazil can shed Giardia cysts that can potentially infect humans, and thus, they may represent a public health risk.     

Fetal infections and antibody profiles in pigs naturally infected with porcine circovirus type 2 (PCV2)

The aim of this study was to describe early infections with porcine circovirus type 2 (PCV2) in naturally infected piglets and the piglets' serologic profiles. A total of 20 sows (15 PCV2-vaccinated and 5 unvaccinated) and 100 newborn piglets were studied. Colostrum and serum of the sows and serum of the presuckling piglets were obtained on the day of parturition. Milk samples were collected on day 20 postpartum. Blood samples were taken and the piglets weighed on days 1, 20, 42, 63, and 84 postpartum. Colostrum and milk were evaluated for infectious PCV2 and for PCV2 total antibody (TA), neutralizing antibody (NA), and IgA. Serum samples were evaluated for PCV2 TA, NA, IgA, IgM, and DNA. The sows had high levels of TA and NA in serum and colostrum; however, 11 and 5, respectively, of the 20 colostrum and milk samples contained infectious PCV2. In the serum, PCV2 DNA and IgM were detected in 17 and 5, respectively, of the 20 sows. Nine piglets were born with PCV2 antibodies, which indicates in utero transmission of PCV2 after the period of immunocompetence (> 70 d of gestation). On day 1 postpartum, PCV2 DNA was detected in 29 of the 100 serum samples from the piglets. There was no difference between the weights of viremic and nonviremic piglets throughout the study. In conclusion, even on farms with sows that have high PCV2 antibody titers, vertical transmission of PCV2 may occur, resulting in piglet infection.     

Comparison of the effects of epidural or intravenous methadone on the minimum alveolar concentration of isoflurane in dogs

The effects of epidural and intravenous (IV) methadone (0.5mg/kg) on the minimum alveolar concentration of isoflurane (ISO(MAC)) were compared in dogs. Six dogs (16.5 ± 2.5 kg bodyweight) received three treatments in random order during isoflurane anaesthesia, with a 7 day washout interval between each study. Methadone was injected via a lumbosacral epidural catheter introduced 10 cm cranially into the epidural canal and the electrical stimulation for ISO(MAC) determination was applied either to the thoracic (EP(T) treatment) or to the pelvic limb (EP(P) treatment) during separate study days. In the IV treatment, ISO(MAC) was determined via electrical stimulation of the pelvic limb. Variables were recorded before (baseline), 2.5 and 5h after drug injection. The ISO(MAC) decreased significantly (P<0.05) from baseline at 2.5 and 5h after methadone in all treatments. At 2.5h, the magnitude of ISO(MAC) reduction did not differ between treatments (mean decreases from baseline: 30-33%). The ISO(MAC) reduction lasted longer following epidural methadone in the thoracic limb (decreases from baseline: 30% at 5h in the EP(T) treatment vs. 19% and 16% in the EP(P) and IV treatments, respectively). Although the isoflurane sparing effect provided by epidural methadone was not significantly greater than IV methadone during the initial stage (2.5h), it was more prolonged than the IV route in specific dermatomes (5h in the thoracic limb) with the epidural technique employed. Methadone may therefore provide a greater isoflurane sparing effect when administered epidurally, compared to IV, when noxious stimulation occurs in specific dermatomes.     

Agreement between direct, oscillometric and Doppler ultrasound blood pressures using three different cuff positions in anesthetized dogs

ObjectiveTo evaluate the agreement between invasive blood pressure (IBP) and Doppler ultrasound blood pressure (DUBP) using three cuff positions and oscillometric blood pressure (OBP) in anesthetized dogs.Study designProspective study.AnimalsNine adult dogs weighing 14.5–29.5 kg.MethodsThe cuff was placed above and below the tarsus, and above the carpus with the DUBP and above the carpus with the OBP monitor. Based on IBP recorded via a dorsal pedal artery catheter, conditions of low, normal, and high systolic arterial pressures [SAP (mmHg) <90, between 90 and 140, and >140, respectively] were induced by changes in isoflurane concentrations and/or dopamine administration. Mean biases ± 2 SD (limits of agreement) were determined.ResultsAt high blood pressures, regardless of cuff position, SAP determinations with the DUBP underestimated invasive SAP values by more than 20 mmHg in most instances. With the DUBP, cuff placement above the tarsus yielded better agreement with invasive SAP during low blood pressures (0.2 ± 16 mmHg). The OBP underestimated SAP during high blood pressures (?42 ± 42 mmHg) and yielded better agreement with IBP for mean (MAP) and diastolic (DAP) arterial pressure measurements [overall bias: 2 ± 15 mmHg (MAP) and 0.2 ± 16 mmHg (DAP)].ConclusionsAgreement of SAP determinations with the DUBP is poor at SAP > 140 mmHg, regardless of cuff placement. Measurement error of the DUBP with the cuff placed above the tarsus is clinically acceptable during low blood pressures. Agreement of MAP and DAP measurements with this OBP monitor compared with IBP was clinically acceptable over a wide pressure range.Clinical relevanceWith the DUBP device, placing the cuff above the tarsus allows reasonable agreement with IBP obtained via dorsal pedal artery catheterization. Only MAP and DAP provide reasonable estimates of direct blood pressure with the OBP monitor evaluated.     

Anti HSV-1 Activity of Halistanol Sulfate and Halistanol Sulfate C Isolated from Brazilian Marine Sponge Petromica citrina (Demospongiae)

The n-butanol fraction (BF) obtained from the crude extract of the marine sponge Petromica citrina, the halistanol-enriched fraction (TSH fraction), and the isolated compounds halistanol sulfate (1) and halistanol sulfate C (2), were evaluated for their inhibitory effects on the replication of the Herpes Simplex Virus type 1 (HSV-1, KOS strain) by the viral plaque number reduction assay. The TSH fraction was the most effective against HSV-1 replication (SI = 15.33), whereas compounds 1 (SI = 2.46) and 2 (SI = 1.95) were less active. The most active fraction and these compounds were also assayed to determine the viral multiplication step(s) upon which they act as well as their potential synergistic effects. The anti-HSV-1 activity detected was mediated by the inhibition of virus attachment and by the penetration into Vero cells, the virucidal effect on virus particles, and by the impairment in levels of ICP27 and gD proteins of HSV-1. In summary, these results suggest that the anti-HSV-1 activity of TSH fraction detected is possibly related to the synergic effects of compounds 1 and 2.     

Spotted Fever Group – Rickettsiae in the Tyrols: Evidence by Seroepidemiology and PCR

The aim of our study was to assess the occurrence of Rickettsia in the inner‐alpine valleys of the Eastern Alps and to determine the amount of seroreaction among the local human population. Ticks were investigated by PCR and the percentage of seropositives was determined among local blood donors by an in‐house immunofluorescence assay. The local cut‐off titre for screening of IgG was set at 1 : 128 with a well‐characterised low‐risk collective according to WHO‐guidelines. Positive sera were confirmed by independent re‐testing. Rickettsia is present in ticks north and south of the continental divide. Of 259 ticks investigated, 12.4% are positive for Rickettsia. Of over 1200 blood donors tested so far, 7.7% bear IgG at a titre of 1 : 128 or higher against R. helvetica. R. helvetica is present in the study area, causes immunoreaction among local residents and is associated with anamnestic erythema. Furthermore, screening with a second Spotted Fever Group Rickettsia indicates that significant parts of the Tyrolean population are exposed to a Rickettsia other than R. helvetica.     

Nephrotoxicity in Goats Caused by Dosing with a Water Extract from the Stems of Narthecium Ossifragum Plants

Seven goats were given a single dose of an aqueous extract derived from 30 g (wet weight) of Narthecium ossifragum per kg liveweight. Their serum creatinine and urea concentrations increased to day 5 but then fell to normal by day 10. Serum magnesium increased to day 4 and decreased to normal by day 9. Their serum calcium concentration was lower than normal on days 4, 5 and 6. Histopathological examination of the kidneys of goats killed or found dead 2, 4, 6, 8, 11 or 16 days after dosing revealed tubular epithelial cell degeneration and necrosis. Regeneration of the tubular epithelium and signs of interstitial fibroplast proliferation and fibrosis could be seen in animals killed on days 8, 11, 16 and 42. No signs of liver damage were observed in 3 goats dosed with the insoluble plant material from 40 g (wet weight) Narthecium ossifragum per kg liveweight. The total dose was divided into three doses, which were given intraruminally within 7 h. The activities of aspartate aminotransferase, -glutamyl-transferase and glutamate dehydrogenase remained within the normal range in all 10 goats after dosing.     

Does the tumour microenvironment alter tumorigenesis and clinical response in transmissible venereal tumour in dogs?

The canine transmissible venereal tumour (CTVT) is a transmissible cancer that is spread naturally between dogs, with the ability to develop and evade the immune system, despite strict immune surveillance of the host. Furthermore, molecular signalling between cells of the immune system and the tumour microenvironment appear to influence the behaviour and development of the tumour. Thus, this study aimed to quantify the expression of genes related to the immune system such as IL‐6, IFN‐γ, and TGF‐β, as well as angiogenic factors (VEGF, CXCR4), in CTVT cells in vivo and in vitro (primary culture), correlating with the clinical response of the animals treated with vincristine. As expected, the most prevalent subtype was plasmacytoid cells, although lymphocytic cells were also found, indicating the possibility of polyclonality. When we compared the gene expressions of IFN‐γ and IL‐6, we mostly found low expression, concluding that MHC expression was probably not occurring in tumour cells, and no activation of immune cells to eliminate the tumour. The TGF‐β gene was normal in the majority of animals but demonstrated decreased expression in vincristine resistant animals, leading to the hypothesis that the concentration of tumour‐derived TGF‐β was affecting and even suppressing the real TGF‐β expression, favouring tumour proliferation and progression in these cases. VEGF expression was extremely high, demonstrating its angiogenic role in tumour growth, while CXCR4 was decreased, possibly because of CTVT’s low metastatic potential. Thus, we concluded that the tumour microenvironment, together with the immune system of the host, influences CTVT, presumably altering its tumorigenesis and the animal’s clinical response to treatment.