Factors affecting urine reagent strip blood results in dogs and nonhuman primates and interpretation of urinalysis in preclinical toxicology studies: a Multi-Institution Contract Research Organization and BioPharmaceutical Company Perspective.

BACKGROUND: Urinalysis data in preclinical toxicology studies can be influenced by preanalytic and analytic factors which have the potential to confound interpretation. There is a paucity of information regarding positive reagent strip urinary blood reactions in healthy nonhuman primates (NHP) and Beagle dogs used in preclinical toxicology studies. OBJECTIVES: The objectives were (1) to establish historical control data for reagent strip urinary blood reactions in healthy NHP and Beagle dogs, (2) to determine the incidence of positive urinary blood reactions during predose and dosing phases, and (3) to determine if collection practice was a relevant parameter. METHODS: Historical control data from 2 institutions in the biopharmaceutical industry were retrospectively analyzed for reagent strip urinary blood reactions in healthy NHP and Beagles. The incidence of positive results between the 2 institutions with different urine collection practices and between males and females was compared. RESULTS: The incidence of positive urinary blood reactions in NHP was comparable between institutions (≤ 14% in males; ≤ 33% in females), while the incidence of positive urinary blood reactions in Beagles was more variable (≤ 77% in males; ≤ 69% in females), and higher in females during the dosing phase. CONCLUSIONS: Positive urinary blood results that could potentially be misinterpreted as toxicologically relevant were identified in healthy NHP and Beagles during predose and dosing phases. Different incidences of positive results between the 2 institutions were likely related to collection practices. Strategies to reduce feces and food contamination of collected urine samples should help minimize false-positive urinary blood reactions.

Avaliação da infecção por leptospiras patogênicas em primatas silvestres mantidos em cativeiro em Salvador, Bahia, Brasil / Evaluation of infection by pathogenic Leptospira in wild primates kept in captivity in Salvador, Bahia, Brazil

Estima-se que 70% das doenças infecciosas emergentes em humanos estão relacionados a exposição a animais silvestres. Estudos envolvendo infecções em primatas não humanos e o seu papel na epidemiologia da leptospirose são escassos.Uma instrução normativa do Ministério do Meio Ambiente(179 de 25/06/2008)recomenda o uso de teste de microaglutinação (MAT) para detecção de anticorpos aglutinantes anti-Leptospira em espécies silvestres em programas de reintrodução à vida livre. Neste contexto, o presente projeto propôs investigar a evidência de prévia exposição e a ocorrência do estado de portador de leptospira patogênicas em primatas silvestres residentes no Parque Zoobotânico Getúlio Vargas de Salvador (PZBGV) e recebidos pelo Centro de Triagem de Animais Silvestres Chico Mendes (CETAS). Foram analisados 42 amostras de soro e sangue total de 42 primatas do PZBGV e 16 amostras de soros de 16 primatas do CETAS pelo teste de aglutinação microscópica (MAT) e PCR respectivamente. Amostras de urina de 3 de primatas do PZBGV e 13 do CETAS também foram submetidas à PCR. A soroprevalência encontrada no PZGV foi de 2%(1/42)enquanto que no CETAS foi de 31%(5/16). O único primata do PZBGV positivo na MAT foi um Allouata caraya (bugio preto) fêmea, adulta, que apresentou reação mista para os sorogrupos Australis e Icterohaemorrhagiae. As cinco amostras positivas do CETAS ocorreram em macacos-prego(Cebus sp.)e foram distribuídos nos sorogrupos: Ballum(1:100), Semaranga (1:200), Grippotyphos (1:100),Cynopeteri(1:100) e uma reação mista Tarassovi / Autumnalis (1:100). Todas as amostras de sangue total e urina analisadas por PCR foram negativas. Conclui-se que a soroprevalência de anticorpos anti-Leptospira foi baixa no PZBGV de Salvador, apesar da alta frequência de roedores na área e endemicidade da leptospirose humana em Salvador. A prevalência elevada foi observada entre os animais resgatados do comércio ilegal no estado da Bahia e esta evidência sorológica de exposição sugere um risco potencial de transmissão da leptospirose ao se adotar estes primatas como animais de estimação

It is estimated that 70% of emerging infectious diseases in humans are related to exposure to wild animals. Studies involving infections in nonhuman primates and their role in the epidemiology of leptospirosis are scarce. A normative statement of the Ministry of the Environment (179 of 25/06/2008) recommends the use of microscopic agglutination test (MAT) for antibodies binding anti-Leptospira in wild species reintroduction programs in the wild. Positive tests indicate the need for quarantine and antimicrobial treatment. In this context, this project proposes to investigate the evidence of prior exposure and the occurrence of carrier status of pathogenic Leptospira in wild primates living in the Parque Vargas Zoobotânico Salvador (PZBGV) and received by the Center for Wildlife Screening Chico Mendes (CETAS). We analyzed 42 serum samples and 42 whole blood PZBGV primates and 16 sera from 16 primates CETAS by microscopic agglutination test (MAT) and PCR respectively. Urine samples of 3 primates PZBGV CETAS and 13 were also subjected to PCR. The seroprevalence in PZBGV was 2% (1/42) while the CETAS was 31% (5/16). The only positive was a primate Allouata caraya (black howler monkey) female, adult, which showed mixed reaction to serogroups Australis and Icterohaemorrhagiae. The five samples positive from CETAS occurred in monkeys (Cebus sp.) And were divided into serogroups: Ballum (1:100), Semaranga (1:200), Grippotyphosa (1:100), Cynopeteri (1:100) and mixed reaction Tarassovi / Autumnalis (1:100). All blood samples and urine samples were analyzed by PCR negative. It is concluded that the seroprevalence of anti- Leptospira antibodies was low in the Zoo of Salvador, despite the high frequency of rodents in the area and endemicity of leptospirosis in Salvador...

[Urine proteins in primates]. / Urinproteine bei Primaten.

Protein excretion and protein fractions either according to their molecular mass or with immunological techniques were studied in the spontaneous morning urine of 17 primate species. The total protein concentration in most of the species ranges between 0.01 and 0.2 mg/ml. The pronounced proteinuria (4 mg/ml) in some south american species (Callithricidae) seems to be remarkable. By using immunoprecipitation (LC-Partigen plates), albumin could be detected in most species, alpha 1-microglobulin in some, and transferrin in few of the species. After electrophoretic separation on pre-cast 1D-micro- SDS-PAA gradient gels (8-25%, semi-automatic Phast-System) followed by CBB R-350 or silver stain respectively, in most species a protein pattern similar to human urine could be observed. As our results show, urine analysis is a suitable tool for noninvasive investigations in primates.

Pregnancy monitoring in diverse primate species by estrogen and bioactive luteinizing hormone determinations in small volumes of urine.

Immunoreactive estrogen and luteinizing hormone/chorionic gonadotropin (LH/CG) bioactivity have been measured throughout pregnancy in diverse primate species with a sample methodological approach utilizing small volumes of urine collected randomly. Total immunoreactive estrogen levels and the relative abundance of component estrogens (estrone, estradiol and estriol) at various stages of gestation were compared in the human, orangutan, pygmy chimpanzee, Douc langur and capuchin. Measurement of urinary LH/CG bioactivity enables an earlier detection of pregnancy when compared to the nonhuman primate pregnancy test. The methods described provide a practical and non-stressful way to detect and monitor pregnancy in widely divergent primate species.

Amylase iso-enzymes in primates of the families Lemuridae, Cebidae, Cercopithecidae, Hylobatidae and Pongidae.

This paper presents the first data in the alpha-amylase iso-enzymes in primates of the families: Lemuridae, Cebidae, Cercopithecidae, Hylobatidae and Pongidae. By means of agar-gel electrophoresis of urine samples from 33 individuals belonging to ten species of the above mentioned families a total of 14 different variants of amylase heterogeneity were found.