Serological analysis was performed to detect morbillivirus infection in Kuril harbor seals in Hokkaido, Japan. Serum samples were collected from the seals at Nosappu (231 sera), Akkeshi (16), and Erimo (75) between 1998 and 2005. Antibodies to phocine distemper virus (PDV) were detected by ELISA in seals from Nosappu and Erimo. Antibodies to PDV were found in 56% (5/9) of the sampled seals from Nosappu in 1998, versus only 5% (3/66) for 2003, 1% (1/79) for 2004, and 1% (1/77) for 2005. These suggest epidemic caused by the virus in or before 1998. As antibody-positive seals included juvenile seals in 2003 and 2005, sporadic infections of the virus are thought to have occurred in recent years. In Erimo, antibodies to PDV were found in 50% (14/28) of sampled seals in 2004, versus only 13% (1/8) for 1999, 7% (1/15) for 2003, and 0% (0/24) for 2005. These suggest sporadic infection by the virus before 2003 and the epizootic between after autumn in 2003, when samples of 2003 were collected, and 2004. Since antibodies to canine distemper virus (CDV) were detected in one adult seal from Nosappu in each year from 2003 to 2005, sporadic infections of the virus, were suggested. There were no difference in incidence of seals with antibodies to the viruses between males and females and between juveniles and adults.

The study determined the types of pesticide/herbicide pollutants in water, sediment and fish from the Kafue River. A preliminary investigation of the oxidative stress from these pesticides/herbicides was also assessed by measurement of catalase activity. Water, sediment and fish samples were collected upstream, midstream and downstream the Kafue river in Chingola, Kitwe, Kafue National Park and Kafue Town. Water, sediment and fish muscle were sampled and analysed for pesticides using Gas chromatography. For catalase activity fish liver samples only were examined. The pesticides/herbicides detected in all samples collectively included: Heptachlor, pp'-DDE, Cypermethrin, Chlordane, Toxaphene, Terbufos, Kelthane, Endosulfan, Dieldrin, pp'-DDD, pp'-DDT, Atrazine, Disulfoton, d-trans-Allethrin and Endrin. On the other hand, catalase activity was detected in all fish liver samples from all sites. Its levels increased significantly from Chingola upstream to sites down-stream with highest being in Kafue town. This study therefore, demonstrates that there is widespread contamination of the Kafue River with pesticides/herbicides. It also demonstrates that organochlorides are found throughout the river especially in fish samples. The spectrum of pesticides/herbicides was much wider in fish probably due to bioaccumulation. It was also observed that fish are subjected to oxidative stress as determined by catalase levels. The stress is more pronounced downstream where the catalase levels were significantly higher than Chingola. The observation that more pesticide varieties are also found downstream may suggest a likely causative effect of the pesticides on oxidative stress although this needs further investigation. This study further tentatively highlights the potential dangers of these agro-related substances to dependants of the Kafue River water body and the need to carry out risk assessments and thereafter institute corrective measures to help reduce contamination and adverse effects.

In genome sequencing project, we encounter the DNA regions that often contain stable secondary structure with high GC content. These regions are difficult to not only amplify by PCR for template preparations, but also deter mine the DNA sequences using standard Cycle sequencing (CS) method. Transcriptional sequencing (TS) is a unique DNA sequencing method using RNA polymerase, and is based on the principles of the chain-termination method, which is a powerful method to analyze GC-rich sequences. In this study, we examined the multiple displacement amplification (MDA) to overcome low efficiency of PCR amplification in GC-rich regions and subjected to TS reaction. Combination of MDA and TS (MDA-TS) was extremely successful with GC content ranging from 65 % to 85%, which are difficult to analyze with PCR and CS. We also report plasmid vector, pTS1, which has the stronger T7 and T3 promoters than those of conventional vectors, and the sequence that decreases transcriptional efficiency was removed from its multiple cloning sites. pTS1 resulted in the improved sequencing accuracy and reduced reaction time up to 5 min. These results showed that MDA-TS is a rapid and accurate method for the analysis of GC-rich templates.

Eco-tourism depending on wildlife is becoming increasingly profitable and landowners are beginning to favor game farming and ecotourism. In these areas, large-scale translocation of wildlife involves a diversity of species and large populations. The African buffalo (Syncerus caffer) is one of the major tourist attractions in Zambia. It accounts for 8.7% and 12.4% of the total animal species hunted in the Game Management Areas and the total hunting revenue earned in Zambia, respectively. It is ecologically an important animal species essential for the purpose of habitat control and facilitating the provision of suitable grazing pastures. However, the rearing of the African buffalo on game ranches has been hampered by its carrier state of the Southern Africa Terroritory (SAT) serotypes of foot and mouth disease virus (FMD). The African buffalo is also known to be a carrier of Theileria parva lawrencei, the causative agent of corridor disease (CD) that continues to have devastating effects on the livestock industry in Zambia. In addition, the importation of buffaloes from countries with populations endemic to bovine tuberculosis is highly restricted. Veterinary regulations in Zambia, strongly advocate against the translocation of buffaloes from protected areas to private ranches for disease control purposes thereby mounting a considerable constraint on the economic and ecological viability of the industry. It is hoped that this review will motivate the relevant government authorities in exploiting ways in which this animal species play a central role in eco-tourism.

Anthrax is endemic in Western and North-western Provinces of Zambia. The disease occurs throughout the year and impacts negatively on the economy of the livestock industry and public health in Zambia. During 1989-1995, there were 1,626 suspected cases of anthrax in cattle in Western province and of these 51 were confirmed. There were 220 cases of human anthrax cases in 1990 alone and 248 cases during 1991-1998 with 19.1% and 7.7% case fatality rates, respectively. Interplay of the ecology of affected areas and anthropogenic factors seem to trigger anthrax epidemics. Anthrax has drawn considerable attention in recent years due to its potential use as a biological weapon. In this paper, the history, current status and approaches towards the control of the disease in Zambia are discussed. Quarantine measures restrict trade of livestock and exchange of animals for draught power resulting in poor food security at household levels. Challenges of anthrax control are complex and comprise of socio-political, economical, environmental and cultural factors. Inadequate funding, lack of innovative disease control strategies and lack of cooperation from stakeholders are the major constraints to the control of the disease.

To develop ELISA to detect antibodies specific to Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), the carboxyl termini of the 34 kDa proteins of M. paratuberculosis and Mycobacterium avium subsp. avium (M. avium) were expressed in Escherichia coli expression system. Antibodies specific to M. paratuberculosis were detected with the truncated 34 kDa protein of M. paratuberculosis in ELISA after pre-absorption of serum samples with the truncated 34 kDa protein of M. avium. All the serum samples from cattle confirmed to be infected with M. paratuberculosis were positive and those from healthy cattle were negative in the present ELISA system. These results indicate that the established ELISA detects antibodies specific to M. paratuberculosis with high specificity and sensitivity and is an useful tool for the screening of Johne's disease.

Genotypes and subgenotypes of bovine viral diarrhea virus (BVDV) field isolates from Japan, Germany and the United States of America (USA) were identified, and the prevalent pattern of BVDV in individual countries was estimated genetically. Subgenotypes were determined based on phylogenetic analyses of nucleotide sequences of a part of the E2-coding gene of BVDV. Forty-five, 61 and 56 BVDV strains were isolated from naturally infected cattle in Japan, Germany and USA, respectively, between 1980 and 2003. The most prevalent BVDV in these three countries was BVDV - 1b. The second most prevalent BVDV strains were 1a, 1d and BVDV - 2 in Japan, Germany and USA, respectively. The most prevalent subgenotype 1b in each country constructed individual small clusters in the subgenotype 1b branch in the phylogenetic tree. Although cattle and/or cattle products were moving among the three countries as part of international trade, the distribution of BVDV in the field in each country showed long-standing individual patterns.

The mitochondrial DNA control region of the sun bear (Helarctos malayanus) was sequenced using 21 DNA samples collected from confiscated sun bears to identify conservation units, such as evolutionarily significant units and management units, in Sarawak, Borneo Island. A total of 10 haplotypes were observed, indicating the presence of at least two lineages in the sun bear population in Sarawak. Presumably, these two lineages could represent evolutionarily significant units. However, the geographical distributions of the two lineages remained unknown due to the lack of information regarding the exact capture locations of the confiscated sun bears. It is essential to elucidate the geographical distributions of these lineages in order to create a proper conservation plan for the sun bears in Sarawak. Therefore, further studies examining the haplotype distributions using DNA samples from known localities are essential.

Em fêmeas bovinas a utilização da técnica de inseminação artificial em tempo fixo (IATF) é possibilitada pelo emprego de fármacos que tem como objetivos sincronizar a emergência das ondas foliculares, os estros e a ovulação. Em rebanhos comerciais, o custo de tais fármacos deve estabelecer uma vantajosa relação com os benefícios. O presente estudo, objetivou comparar as taxas de prenhez em vacas Nelore (Bos taurus indicus) inseminadas em tempo fixo, tratadas com implantes auriculares contendo 3mg de norgestomet (Crestar®), novos (IN) ou utilizados uma vez (IR; reutilizados), associados a administração de norgestomet (NG) e valerato de estradiol (VE) ou progesterona (P4) e benzoato de estradiol (BE). Vacas Nelore PO (n=241), amamentando, com o bezerro ao pé, receberam um dos quatro tratamentos: Crestar®novo durante 10 dias associado a administração de 3mg de NG e 5mg de VE (grupo IN/NG+VE; n=61); Crestar®novo inserido durante 8 dias associado a 50mg de P4 e 2mg de BE (grupo IN/P4+BE; n=61); Crestar®reutilizado inserido durante 10 dias associado a administração de 3mg de NG e 5mg de VE (grupo IR/NG+VE; n=58) ou Crestar®reutilizado inserido durante 8 dias associado a e 50mg de P4 e 2mg de BE (grupo IR/P4+BE; n=61). No dia da remoção dos implantes os animais receberam 7,5mg de Luprostiol e 24 horas após a remoção 1mg de BE. A IATF foi realizada 54 a 56 horas após a retirada dos implantes. Após a IATF, as vacas tiveram os estros observados durante um período de 49 dias e em estro foram reinseminadas 12 horas após a observação. O diagnóstico de gestação foi realizado por ultra-sonografia 35 dias após a IATF e após o final da estação de monta. Foram avaliadas as taxas de prenhez na IATF (TP IATF) e no final da estação de monta (TP EM). Não houve interação entre as características dos implantes (novos e reutilizados) e os tratamentos administrados no dia da inserção do implante (NG+VE e P4+BE). A utilização do CIDR®novo ou reutilizado não teve efeito nas TP IATF (48,3% vs 48,7%) e nas TP EM (85,2 vs 86,5%). Os tratamentos com NG+VE e P4+BE não tiveram efeito nas TP IATF (49,5 vs 47,5%) e TP EM (86,5 vs 85,2%). Houve efeito do número de parições (primíparas e multíparas) nas TP IATF (35% vs. 52,7%; P<0,01) e nas TP EM (71,9% vs. 90,2%; P<0,01). Conclui-se que implantes de norgestomet novos e reutilizados, quando associados ao NG+VE ou P4+BE promovem taxas de prenhez bastante satisfatórias em vacas Nelore e que melhores taxas são obtidas em fêmeas multíparas. | The use of fixed-time artificial insemination (IATF) in cows is possible by the use of drugs that aim at the synchronization of the folicular wave pool, the estrus and ovulation. In trading cattle the costs of these drugs must stablish a profitable relation with the benefits. The present study meant to compare the pregnancy rates in Nelore (Bos taurus indicus) cows inseminated at fixed-time, treated with new (IN) or once used (IR; reused) auricular releasing devices with 3mg of norgestomet (Crestar®), associated with the administration of norgestomet (NG) and estradiol valerate (VE) or progesterone (P4) and estradiol benzoate (BE). Pure breed Nelore cows (n=241), on lactation with calf received one of the four treatments: new Crestar®during 10 days administrated with 3mg of NG and 5 mg of VE (group IN/NG+VE; n=61); new Crestar®inserted during 8 days with 50mg of P4 and 2mg of BE (group IN/P4+BE; n=61); reused Crestar®inserted during 10 days in association with 3 mg of norgestomet and 5 mg of VE (group IR/NG+VE; n=58) or reused Crestar®inserted during 8 days in association with 50mg of P4 and 2mg of BE (group IR/P4+BE; n=61). On the day the releasing devices were removed the animals received 7,5mg of Luprostiol and after 24 hours 1mg of BE. A fixed-time artificial insemination was done 54 to 56 hours after the removal of the releasing devices. Cows detected in estrus after the insemination at fixed-time were observed during a period of 49 days and reinseminated. The pregnancy diagnosis was done by ultrassonography 35 days after the IATF and after the end of the breeding season. The pregnancy rates of IATF (TP IATF) and at the end of the breeding season (TP EM) were estimated. There was no interaction between the characteristics of the (new or reused) releasing devices and the treatment given at the day the releasing devices were inserted (NG+VE e P4+BE). The use of new or used CIDR®had no effect on TP IATF (48,3% vs 48,7%) and on TP EM (85,2% vs 86,5%). The treatments with NG+VE e P4+BE did not have effect on TP IATF (49,5% vs 47,5%) and TP EM (86,5 vs 85,2%). There was a effect on the number of parturition (primipara and multipara) on the TP IATF (35% vs 52,7%; P<0,01) and on TP EM (71,9% vs 90,2%; P<0,01). It can be concluded that the new or once used releasing devices with norgestomet when associated with NG+VE or P4+BE promote highly satisfatory pregnancy rates in Nelore cows, being the better rates obtained in multipara females.

Para descrever a anatomia ultra-sonográfica foram examinados os ligamentos patelares (medial, intermédio e lateral) e as estruturas relacionadas de 20 articulações fêmuro-tibio-patelares de cavalos adultos. Tanto a ecogenicidade como o alinhamento das fibras de colágeno, a espessura, a forma e relação com as estruturas vizinhas foram avaliadas. Os três ligamentos apresentaram ecogenicidade uniforme e paralelismo das fibras colágenas. Em secção longitudinal os ligamentos patelares medial (LPM) e intermédio (LPI) apresentaram maior espessura do segmento distal (p<0,01) quando comparado com o proximal. Comparando-se o mesmo segmento entre os três ligamentos, o LPI apresentou espessura maior (p<0,01) em relação ao LPM e ligamento patelar lateral (LPL). Em secção transversal (cm²) o LPL apresentou a maior área (p <0,01), seguido pelo LPI e LPM. Em secção transversal do segmento médio, o LPM apresentou forma triangular, o LPI em forma triangular ou circular, e o LPL formato achatado. As estruturas adjacentes identificadas durante o exame dos ligamentos foram cristas femorais, cartilagem articular, tecido gorduroso periligamentar, patela e tíbia proximal. | In order to describe the ultrasonographic anatomy of patellar ligaments (medial, middle, lateral) and associated structures, 20 femorotibiopatellar joints of adult horses were investigated. Echogenicity, collagen fibers alignment, size, shape and surrounding structures were evaluated. All ligaments presented uniform echogenicity and collagen fibers parallelism. The longitudinal section of the medial and middle patellar ligaments presented a significant larger thickness (p<0,01) of the distal region when compared to the proximal region. When comparing the same site between the three ligaments, LPI was thicker (p<0,01) than lateral patellar ligament (LPL) and LPM. The LPL transversal section (cm²) presented a significant larger area (p<0,01), followed by the LPI and LPM. The transversal section of the medium region revealed a LPM with triangular shape, triangular or circular LPI, and flattened LPL. The surrounding structures observed during the evaluation were: femoral groove, articular cartilages, periligamentar fat tissue, patella and proximal tibia.