Cloning, expression and characterization of monkey (Macaca fascicularis) CD137

CD137 plays an important role as a co-stimulatory molecule in activated T cells. Agonistic CD137 specific antibodies have been investigated as therapeutic agents to promote tumor-specific immune responses by direct activation of T cells. As part of the pre-clinical pharmacological evaluation of cynomolgus monkeys, monkey CD137 was cloned and characterized. The deduced amino acid sequence encoded a full-length gene of 254 amino acids 95% identical to human CD137. Sequence variants identified in monkey CD137 include four splice variants lacking the transmembrane domain. These variants were detectable in human including two previously unreported variants. Two missense single nucleotide polymorphisms were detected present in 42 and 50% of 36 monkeys tested. In both monkey and human, mRNA expression of full-length CD137 and splice variants were significantly increased in peripheral blood mononuclear cells (PBMCs) upon stimulation by anti-CD3 antibodies. Recombinant monkey CD137 protein was bound with high affinity by an agonistic anti-human CD137 antibody but not by an anti-mouse CD137 antibody. In summary, compared to human, monkey CD137 showed distinct extracellular domain amino acid sequence and sequence polymorphisms. Thus, antibodies directed against epitopes in this extracellular domain could have differences in pharmacologic activity between cynomolgus monkeys and human or across individual cynomolgus monkeys.     

Identification of unreported putative new bovine papillomavirus types in Brazilian cattle herds

The amplification by degenerate primers FAP59/FAP64 and sequencing allowed the detection of 15 putative new BPV types in cutaneous warts as well as in healthy skin. Four of these isolates were recently recognized as new BPV types (BPV-7, -8, -9, and -10) after determination of their complete genome sequences. In Brazil, investigations involving the definition of BPV types present in skin warts are still rare. The aim of the current study was to identify the BPV types associated with cutaneous papillomatosis observed in Brazilian cattle herds. Twenty-two cutaneous papilloma specimens were submitted to PCR assay employing the FAP primer pair. All PCR products with approximately 480 bp were submitted to direct sequencing. Cloning was performed for the amplicons which prior analysis revealed as putative new BPV types. From 16 cutaneous lesions, BPV-1, -2, and -6 were identified in two, six, and eight papilloma specimens, respectively. In addition, four putative new BPV types were identified in other six skin warts, and then designated as BPV/BR-UEL2 to -5. The detection of the BPV-1, -2, and -6 types in skin wart specimens supports the existence of these BPV types throughout the Brazilian cattle herd. In addition, the identification of four putative new BPV types is the first report of the presence of different BPV types in the American continent.     

Post mortem parasitological evaluation of dogs seroreactive for Leishmania from Rio de Janeiro, Brazil

A parasitological study was conducted on 66 dogs seroreactive for Leishmania captured as a control measure of visceral leishmaniasis in the State of Rio de Janeiro, Brazil. Biological samples from different anatomical sites were collected during autopsy of the animals and cultured on biphasic medium (NNN/Schneider). The Leishmania isolates were characterized by isoenzyme electrophoresis. Leishmania was isolated from 80.3% of the animals: 12 animals with Leishmania (Viannia) braziliensis isolated exclusively from cutaneous lesions, 39 with L. (L.) chagasi isolated from different sites in the same animal, and 2 with simultaneous isolation of L. (V.) braziliensis from cutaneous lesions and L. (L.) chagasi from different sites. Isolation in culture revealed the absence of Leishmania parasites in 13 animals. The results obtained confirm the existence of mixed infections in dogs in Rio de Janeiro and indicate the need to complement the investigation of seroreactive dogs using methods for the parasitological diagnosis and identification of Leishmania species.     

Where do we go in protection against pathogens?

A few reflections are made on the future strategies and approaches for combating infectious diseases of livestock, keeping into consideration public concern on food quality and availability. The policy of eradication of pathogens/diseases instead of vaccination has rendered livestock naive to certain pathogens and as a consequence susceptible to epidemic outbreaks, even more so as we have to conclude that outside attack is quite often difficult to control. Therefore, immunoprophylactic measures in livestock remain top priority and will in the future focus more on the induction of mucosal immunity and the activation of innate immunity.     

Use of the scrotum as a transposition flap for closure of surgical wounds in three dogs

OBJECTIVE: To describe use of the scrotum as a pedicle flap to cover defects created by tumor excision in the perineum or caudal and medial aspect of the thigh. STUDY DESIGN: Clinical study. ANIMALS: Three dogs. METHODS: After tumor excision and prescrotal castration, an incision was made around the base of the scrotum, leaving a pedicle on the side opposite the skin defect. After removal of the tunica dartos and abdominal fascia, the scrotal skin was stretched and shaped to the defect, then sutured in position with a simple interrupted pattern. Bandages that were changed every 48 hours compressed the flaps, and healing was observed until sutures were removed. RESULTS: Two dogs had first intention healing in 15 days, whereas 1 dog developed necrosis of 10% of the flap, and this area healed by second intention. CONCLUSION: In male dogs, the scrotal skin can be used as a pedicle flap for reconstructive surgery of wounds in the perineum and the proximomedial and caudal aspect of the thigh. CLINICAL RELEVANCE: The perineal region remains a surgical challenge because of the lack of the available skin for reconstruction of surgical wounds. The scrotal skin should be considered for use as a transposition flap to cover skin defects in this region.     

Haptoglobin and pig-major acute protein are increased in pigs with postweaning multisystemic wasting syndrome (PMWS)

The objective of this study was to determine the serum concentration levels of selected acute phase proteins (APP), haptoglobin (HPT) and pig-major acute phase protein (pig-MAP), in postweaning multisystemic wasting syndrome (PMWS) affected pigs and PCV2-subclinically infected pigs. In a first study, a group of 15 eight-week-old conventional pigs from a PMWS affected farm were bled and a complete necropsy, histopathology and in situ hybridisation to detect PCV2 were performed. Based on the results, pigs were classified as suffering from PMWS (n = 10) or healthy animals (n = 5). In a second study, a group of 45 pigs from another PMWS affected farm were selected and bled at 3, 7, 12 and 28 weeks of age. The assessment of PCV2 infection status in these pigs was retrospectively done by PCV2 PCR in serum samples. Selected APP were measured in the serum of all studied pigs by means of radial immunodiffusion. Mean HPT and pig-MAP levels were significantly increased (p = 0.004 and p = 0.0006 respectively) in PMWS-affected pigs when compared to levels found in healthy pigs (2.52 +/- 0.88 mg/mL vs. 1.06 +/- 0.73 mg/mL for HPT and 3.81 +/- 1.53 mg/mL vs. 0.76 +/- 0.34 mg/mL for pig-MAP). In the second study, no significant difference in mean HPT and pig-MAP values were observed between PCV2 PCR positive and negative pigs of any age. However, both APP increased significantly with age in PCV2 PCR negative pigs. Altogether, the present results suggest that APP levels are significantly increased in pigs that develop PMWS, but not in animals with a PCV2 subclinical infection.     

Multidrug-Resistant Pseudomonas aeruginosa Isolates From Captive Reptiles

Pseudomonas aeruginosa represents an opportunistic pathogen for animals and humans that is often associated with high disease morbidity and, at times, mortality. Captive reptiles have been shown to be reservoirs of P. aeruginosa strains that can be sources of exposure to humans that come in contact with these animals. In this study, the prevalence of P. aeruginosa among subclinical captive reptile species and the antimicrobial sensitivity of bacterial isolates were investigated. Sixty-five oral swabs were collected from captive reptiles belonging to 15 different species in which no overt signs of disease were evident. From this group of animals, 46 (70.8%) isolates were identified as P. aeruginosa. All of the P. aeruginosa strains were shown to have a wide range of antibiotic resistance. At present, there is a paucity of data regarding the prevalence of P. aeruginosa in various reptile species; therefore, continued scientific investigations are indicated to determine the significance of P. aeruginosa infection as it relates to captive reptile species.     

Developmental competence of heat stressed oocytes from Holstein and Limousine cows matured in vitro

The negative effects of heat stress on dairy cattle's fertility have been extensively studied, but the relevant knowledge for beef cattle is rather limited. The aims of this study were to investigate the effects of HS during in vitro maturation on the developmental potential of oocytes derived from Limousine and Holstein cows and to estimate the effect of the differential gene expression of important genes in oocytes, cumulus cells and blastocysts in the growth competence between the breeds. In seven replicates, cumulus oocyte complexes from Holstein and Limousine cows were matured for 24 hr at 39°C (controls C; Hol_39, Lim_39) or at 41°C from hour 2 to hour 8 of IVM (treated T; Hol_41, Lim_41), fertilized, and presumptive zygotes were cultured for 9 days at 39°C. Cleavage and embryo formation rates were evaluated 48 hr post-insemination and on days 7, 8 and 9, respectively. From all groups, subsets of cumulus cells, oocytes and blastocysts were analysed for the relative expression of genes related to metabolism, stress, apoptosis and placentation. No difference was detected in cleavage rate or in blastocyst formation rate among the control groups. In both breeds, heat stress reduced blastocyst yield, but at all days the suppression was higher in Limousines. In Holsteins, altered gene expression was detected in cumulus cells (G6PD, GLUT1) and blastocysts (PLAC8), while in Limousines, differences were found in oocytes (G6PD, HSP90AA1), in cumulus cells (CPT1B, HSP90AA1, SOD2) and blastocysts (DNMT, HSP90AA1, SOD2). It appears that Holstein COCs are more tolerant than Limousine COCs, possibly due to compulsory, production driven selection.