A light-touch routing optimization tool (RoOT) for vaccine and medical supply distribution in Mozambique.

Planning vaccine distribution in rural and urban poor communities is challenging, due in part to inadequate vehicles, limited cold storage, road availability, and weather conditions. The University of Washington and VillageReach jointly developed and tested a user-friendly, Excel spreadsheet based optimization tool for routing and scheduling to efficiently distribute vaccines and other medical commodities to health centers across Mozambique. This paper describes the tool and the process used to define the problem and obtain feedback from users during the development. The distribution and routing tool, named route optimization tool (RoOT), uses an indexing algorithm to optimize the routes under constrained resources. Numerical results are presented using five datasets, three realistic and two artificial datasets. RoOT can be used in routine or emergency situations, and may be easily adapted to include other products, regions, or logistic problems.

Retrospective and prospective studies of transmissible viral proventriculitis in broiler chickens in Brazil.

We investigated the occurrence and pathologic findings of transmissible viral proventriculitis (TVP) associated with the chicken proventricular necrosis virus (CPNV) in commercial broiler chickens in southeastern Brazil. Seventy-three broilers, 25-36 d old, with a history of reduced growth, were referred to our veterinary pathology services from 2013 to 2017. Broilers were clinically examined, weighed, and euthanized for postmortem examination. Broilers of different ages with proventricular histologic lesions were positive for CPNV by RT-PCR; however, the intensity of histologic lesions was higher among 33-d-old animals, and viral RNA detection was more frequent among those that were 28 d old. In the proventriculi of 35 of 73 (48%) broilers, lesions were characterized by glandular epithelial necrosis, lymphoplasmacytic and histiocytic infiltrates, and metaplasia of glandular epithelium to ductal epithelium. In 24 of 73 (36%) broilers with histologic TVP-compatible lesions, CPNV was detected by RT-PCR for the viral protein 1 (VP1) gene. Broilers with histologic lesions were lighter than expected compared to the Cobb 500 standard weight. TVP has not been reported previously in broiler chickens in Brazil, to our knowledge.

Feline coronavirus isolates from a part of Brazil: insights into molecular epidemiology and phylogeny inferred from the 7b gene.

The Feline coronavirus (FCoV) can lead to Feline infectious peritonitis (FIP), which the precise cause is still unknown. The theory of internal mutation suggests that a less virulent biotype of FCoV (FECV) would lead to another more pathogenic biotype (FIPV) capable of causing FIP. In this work, the 7b gene was amplified from 51 domestic cat plasma samples by semi-nested PCR and tested through phylogenetic and phylogeographical approaches. The 7b gene of Brazilian isolates displayed high conservation, a strong correlation between the geographic origin of the viral isolates and their genealogy, and its evolution was possibly shaped by a combination of high rates of nucleotide substitution and purifying selection.

Fatal necrotic tracheitis by Aviadenovirus in captive Alagoas curassows (Pauxi mitu) extinct from the wild.

Extinct from nature, captive young Alagoas curassows (Pauxi mitu) were found agonizing or dead with respiratory disease. Intranuclear inclusion bodies were found in the epithelia of the trachea, associated with marked necrotic tracheitis. An Aviadenovirus was isolated in chicken eggs and characterized genetically with 99% identity to the fowl Aviadenovirus A, as based on the hexon protein gene. This is the first report of respiratory disease caused by Aviadenovirus in any cracid species in Brazil, recommending for stricter biosecurity in the conservation premises. RESEARCH HIGHLIGHTS Fatal tracheitis in curassows extinct from nature was associated with Aviadenovirus A. Seven-month-old Alagoas curassows (Aves: Cracidae) died with haemorrhagic tracheitis. Aviadenovirus A with 99% identity to fowl adenovirus 1 was detected in dead curassows. Fatal tracheitis by Aviadenovirus was described in Pauxi mitu (Aves: Cracidae).

Avian Pox in Native Captive Psittacines, Brazil, 2015.

To investigate an outbreak of avian pox in psittacines in a conservation facility, we examined 94 birds of 10 psittacine species, including sick and healthy birds. We found psittacine pox virus in 23 of 27 sick birds and 4 of 67 healthy birds. Further characterization is needed for these isolates.

Genotypic characterization of psittacid herpesvirus isolates from Brazil

Abstract Thirty-six isolates of psittacid herpesvirus (PsHV), obtained from 12 different species of psittacids in Brazil, were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis and PCR amplification. RFLP analysis with the PstI enzyme revealed four distinct restriction patterns (A1, X, W and Y), of which only A1 (corresponding to PsHV-1) had previously been described. To study PCR amplification patterns, six pairs of primers were used. Using this method, six variants were identified, of which, variants 10, 8, and 9 (in this order) were most prevalent, followed by variants 1, 4, and 5. It was not possible to correlate the PCR and RFLP patterns. Twenty-nine of the 36 isolates were shown to contain a 419 bp fragment of the UL16 gene, displaying high similarity to the PsHV-1 sequences available in GenBank. Comparison of the results with the literature data suggests that the 36 Brazilian isolates from this study belong to genotype 1 and serotype 1.

Genotypic characterization of psittacid herpesvirus isolates from Brazil.

Thirty-six isolates of psittacid herpesvirus (PsHV), obtained from 12 different species of psittacids in Brazil, were genotypically characterized by restriction fragment length polymorphism (RFLP) analysis and PCR amplification. RFLP analysis with the PstI enzyme revealed four distinct restriction patterns (A1, X, W and Y), of which only A1 (corresponding to PsHV-1) had previously been described. To study PCR amplification patterns, six pairs of primers were used. Using this method, six variants were identified, of which, variants 10, 8, and 9 (in this order) were most prevalent, followed by variants 1, 4, and 5. It was not possible to correlate the PCR and RFLP patterns. Twenty-nine of the 36 isolates were shown to contain a 419bp fragment of the UL16 gene, displaying high similarity to the PsHV-1 sequences available in GenBank. Comparison of the results with the literature data suggests that the 36 Brazilian isolates from this study belong to genotype 1 and serotype 1.

Cirurgia oncoplástica e reconstrutiva da mama: Reunião de Consenso da Sociedade Brasileira de Mastologia / Oncoplastic and reconstructive surgery of the breast: Consensus Meeting of the Brazilian Society of Mastology

Objetivo: Apesar do grande número de publicações em cirurgia oncoplástica e reconstrutiva da mama, diversas questões permanecem controversas. Assim, o objetivo desta Reunião de Consenso, foi desenvolver um guia prático de recomendações baseadas nas melhores evidências disponíveis na literatura. Método: Os painelistas foram os membros da Comissão de Cirurgia Oncoplástica e Reconstrutiva da Sociedade Brasileira de Mastologia. A reunião foi realizada em agosto de 2015 em Bento Gonçalves (RS). Cada painelista recebeu e respondeu previamente um questionário com 46 itens, com base na melhor evidencia cientifica e em sua experiência. Foi considerado consenso a concordância de 75% entre painelistas. Resultados: Houve consenso em 25 itens, dos quais para oito houve concordância de 100%. O mais importantes foram: comprometimento das margens em cirurgia oncoplástica pode ser resolvido com ampliação de margens na maioria dos casos; tumores multifocais não são contraindicação para cirurgia oncoplástica; idade >70 anos não representa contraindicação para uso de técnicas oncoplásticas; reconstrução imediata pode ser indicada com segurança para a maioria das candidatas à mastectomia; pacientes com indicação de radioterapia pós-mastectomia podem ser submetidas à reconstrução imediata, devendo ter ciência dos riscos maiores para mau resultado estético; mastectomia com preservação do complexo areolopapilar é segura nos casos de câncer; radioterapia após a mastectomia com preservação do complexo areolopapilar não está indicada fora dos critérios clássicos de irradiação do plastrão; tela abdominal reduz chances de hernia no caso de reconstrução com TRAM. Conclusão: através desta reunião foi possível estabelecer importantes pontos consensuais de acordo com a opinião dos especialistas, que poderão auxiliar os mastologistas na tomada de decisões em cirurgias oncoplásticas e reconstrutivas da mama.

Objective: Despite the large number of publications in oncoplastic and breast reconstructive surgery, several issues remain controversial. The aim of this Consensus Meeting was to develop a practical guide of recommendations based on the best evidence in the literature. Method: All panelists were members of the Oncoplastic Commission of the Brazilian Society of Mastology. The Consensus Meeting was held in Bento Gonçalves (RS), in August 2015. Each panelist received and answered a questionnaire with 46 items, based on the best evidence in the literature and in their expertise. It was considered consensus the agreement of 75% between panelists. Results: There was consensus on 25 items, of which eight were for 100% agreement. The most important of these topics were: involvement of the margins in oncoplastic surgery can be solved by resection of margins in most cases; multifocal tumors is not a contraindication for oncoplastic surgery; age >70 years is not a contraindication for use of oncoplastic techniques; immediate reconstruction can be performed safely to most candidates for mastectomy; patients for post-mastectomy radiotherapy may be subject to immediate reconstruction and should be aware of the risks for poor aesthetic result; mastectomy with preservation of the nipple and areola complex is safe in cancer; radiotherapy after mastectomy with preservation of the nipple and areola complex is not indicated outside the classical criteria of irradiation chest wall; abdominal mash reduces chances of hernia in TRAM flaps. Conclusion: In this meeting it was possible to establish important consensus points according to the opinion of experts, which can help breast surgeons in their decision-making in oncoplastic and reconstructive surgery of the breast.

Biased Random-Key Genetic Algorithms for the Winner Determination Problem in Combinatorial Auctions.

In this paper we address the problem of picking a subset of bids in a general combinatorial auction so as to maximize the overall profit using the first-price model. This winner determination problem assumes that a single bidding round is held to determine both the winners and prices to be paid. We introduce six variants of biased random-key genetic algorithms for this problem. Three of them use a novel initialization technique that makes use of solutions of intermediate linear programming relaxations of an exact mixed integer linear programming model as initial chromosomes of the population. An experimental evaluation compares the effectiveness of the proposed algorithms with the standard mixed linear integer programming formulation, a specialized exact algorithm, and the best-performing heuristics proposed for this problem. The proposed algorithms are competitive and offer strong results, mainly for large-scale auctions.

Fatal proventricular dilatation disease in captive native psittacines in Brazil.

An outbreak of proventricular dilatation disease (PDD), a fatal inflammatory disease of psittacines (Aves: Psittaciformes), is described in native Brazilian psittacines. Twenty captive psittacines that died of suspected PDD were necropsied and 10 were submitted to histopathology, reverse transcriptase PCR (RT-PCR), and immunohistochemistry (IHC) for avian bornavirus (ABV). Examined species were one pileated parrot (Pionopsitta pileata), three vinaceous-breasted parrots (Amazona vinacea), two blue-winged macaws (Primolius maracana), one scarlet macaw (Ara macao), one chestnut-fronted macaw (Ara severa), one scaly-headed parrot (Pionus maximiliani), and one red-browed Amazon parrot (Amazona rhodocorytha). Gross examination and histopathology revealed typical PDD lesions in all birds. The presence of ABV was confirmed in four psittacines including one red-browed Amazon parrot, one blue-winged macaw, one scarlet macaw, and one chestnut-fronted macaw. In the red-browed Amazon parrot and in one blue-winged macaw, IHC demonstrated ABV antigens in the nucleus and cytoplasm of cells in various organs. This is the first description of PDD by ABV in Brazilian psittacines and indicates the necessity for adopting a strategic control plan for reducing its impact in native birds.

On the improvement of blood sample collection at clinical laboratories.

BACKGROUND: Blood samples are usually collected daily from different collection points, such hospitals and health centers, and transported to a core laboratory for testing. This paper presents a project to improve the collection routes of two of the largest clinical laboratories in Spain. These routes must be designed in a cost-efficient manner while satisfying two important constraints: (i) two-hour time windows between collection and delivery, and (ii) vehicle capacity. METHODS: A heuristic method based on a genetic algorithm has been designed to solve the problem of blood sample collection. The user enters the following information for each collection point: postal address, average collecting time, and average demand (in thermal containers). After implementing the algorithm using C programming, this is run and, in few seconds, it obtains optimal (or near-optimal) collection routes that specify the collection sequence for each vehicle. Different scenarios using various types of vehicles have been considered. Unless new collection points are added or problem parameters are changed substantially, routes need to be designed only once. RESULTS: The two laboratories in this study previously planned routes manually for 43 and 74 collection points, respectively. These routes were covered by an external carrier company. With the implementation of this algorithm, the number of routes could be reduced from ten to seven in one laboratory and from twelve to nine in the other, which represents significant annual savings in transportation costs. CONCLUSIONS: The algorithm presented can be easily implemented in other laboratories that face this type of problem, and it is particularly interesting and useful as the number of collection points increases. The method designs blood collection routes with reduced costs that meet the time and capacity constraints of the problem.

Outbreak of human herpesvirus type 1 infection in nonhuman primates (Callithrix penincillata).

Human herpesvirus type 1 (HHV-1) is widely dispersed among the human population. Although infection is often asymptomatic in humans, nonhuman primates develop a severe and often fatal infection. In August 2006, 13 black-tufted marmosets (Callithrix penincillata) from a group of 14 presented with clinical apathy, anorexia, and ataxia. Physical examination revealed conjunctivitis, erosive or ulcerative lesions on the skin, and swollen lymph nodes. Of the 14 animals captured, 10 died. Grossly, ulcers and erosions were observed on the skin of face, nasal planum, lips, and oral mucosa. Histologically, superficial vesicular and erosive stomatitis with associated basophilic intranuclear inclusion bodies in the squamous epithelium were observed. Swabs from oral lesions and tissue samples from necropsied animals were positive for HHV-1 by nested polymerase chain reaction for eight animals.

Identification and isolation of psittacid herpesvirus from psittacids in Brazil.

Psittacid herpesvirus (PsHV) was isolated from 41 birds kept in captivity in Belo Horizonte, Minas Gerais/Brazil using chicken embryo fibroblasts (CEF) cell cultures. For this study, leukocytes or cloacal swabs of live birds were used. Also, portions of liver, spleen or kidney from birds collected at necropsy were utilized for these tests. PCR tests confirmed the presence of PsHV in 100% of samples. Thirty-three of the PCR products were sequenced and the results disclosed a 99% and 100% identity when compared with other sequences PsHV-1 (AY372243.1 and AF261756.1), previously deposited in GenBank. In addition, histopathology was performed and 19 of the 29 birds contained random multifocal lymphoplasmacytic hepatitis with necrotic foci, suggestive of viral infection. Three samples were examined by electron microscopy to visualize the viral particles obtained from cell culture. The viral structures measured 269 nm in average, had envelopes with an icosahedral capsid and tegument, consistent with herpesvirus. Thus, a total of 41 isolates were obtained from PsHV cell cultivation in CEF, confirming the circulation of the virus between parrots kept in captivity in Belo Horizonte, and affirming the importance of further studies in this area.

The paradox of feline coronavirus pathogenesis: a review.

Feline coronavirus (FCoV) is an enveloped single-stranded RNA virus, of the family Coronaviridae and the order Nidovirales. FCoV is an important pathogen of wild and domestic cats and can cause a mild or apparently symptomless enteric infection, especially in kittens. FCoV is also associated with a lethal, systemic disease known as feline infectious peritonitis (FIP). Although the precise cause of FIP pathogenesis remains unclear, some hypotheses have been suggested. In this review we present results from different FCoV studies and attempt to elucidate existing theories on the pathogenesis of FCoV infection.

Isolation and characterization of gingival fibroblasts positive for alkaline phosphatase in patients with chronic periodontitis and drug-induced gingival hyperplasia / Isolamento e caracterização de fibroblastos gengivais positivos para fosfatase alcalina de pacientes portadores de periodontite crônica e hiperplasia gengival medicamentosa

Purpose: To investigate the expression of alkaline phosphatase (ALP) activity in gingival fibroblasts from individuals with chronic periodontitis (CP) and drug-induced gingival hyperplasia (DGH) induced by diphenylhydantoin. Methods: Gingival fragments were obtained from 13 patients (8 women and 5 men, from 22 to 74 years of age), with 4 fragments from clinically normal gingiva (NG), 5 from biopsy of periodontal pockets with CP, and 4 from DGH induced by diphenylhydantoin. Using an enzymatic digestion procedure, gingival cell suspensions containing ALP-positive fibroblasts were prepared without affecting ALP activity. Cytochemistry and histochemistry analyses were performed. Results: Fibroblasts from NG presented low levels of ALP when compared to CP and DGH, which showed elevated and intermediate levels of ALP, respectively. Little cell proliferation was observed for fibroblasts from CP and DGH as compared to NG. However, the quantity of cells recovered from the subcultures was similar to the quantity recovered from the initial cell culture for the three sources. Conclusion: The expression of ALP is increased in CP and DGH, and fibroblasts in CP and DGH show low proliferation. This suggests that periodontal inflammation and diphenylhydantoin may influence ALP expression and human gingival fibroblast expansion. Other studies are necessary to better assess the importance of ALP in the development and progression of CP and DGH.

Objetivo: Investigar a expressão da atividade de fosfatase alcalina (ALP) em fibroblastos gengivais de sujeitos com periodontite crônica (PC) e hiperplasia gengival medicamentosa (HGM) induzida por difenilhidantoína. Metodologia: Os fragmentos gengivais foram obtidos de 13 pacientes (8 mulheres e 5 homens, 22 a 74 anos), sendo 4 fragmentos procedentes de gengiva clinicamente normal (GN), 5 de biópsias de bolsas periodontais com PC e 4 de HGM induzida pela difenilhidantoína. Através de digestão enzimática, suspensões de células gengivais foram preparadas contendo fibroblastos ALP-positivos sem afetar a atividade da enzima ALP. Análises citoquímica e histoquímica foram realizadas. Resultados: Fibroblastos de GN apresentaram fraca positividade para ALP quando comparados com PC e HGM. PC e HGM exibiram níveis elevados e intermediários, respectivamente, de ALP. Observou-se menor proliferação celular na cultura de células em PC e HGM que em GN. Entretanto, a quantidade de células recuperadas das sub-culturas foi similar à quantidade recuperada na cultura inicial para as três linhagens. Conclusões: A expressão de ALP na PC e na HGM encontra-se aumentada e ocorre baixa proliferação de fibroblastos na PC e na HGM. Isto sugere que a inflamação periodontal e a difenilhidantoína podem influenciar a expressão de ALP e a multiplicação de fibroblastos gengivais humanos. Outros estudos são necessários para melhor avaliar a importância de ALP no desenvolvimento e na progressão de PC e HGM.

Survey of feline leukemia virus and feline coronaviruses in captive neotropical wild felids from Southern Brazil.

A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center (Refúgio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44 animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas reverse transcriptase-PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these species in Southern Brazil.

An outbreak of malignant catarrhal fever in Murrah buffaloes in Minas Gerais, Brazil / Surto de febre catarral maligna (FCM) em búfalos da raça Murrah no estado de Minas Gerais

An outbreak of Malignant Catarrhal Fever (MCF) resulted in death of five female buffaloes and one domestic cow from the same farm. Four buffaloes died 10-15 days after the appearance of clinical signs, while the fifth was euthanized in extremis, after similar clinical signs. Histopathological lesions included multifocal histiolymphocytic epicarditis, myocarditis and lymphocytic interstitial pneumonia, which are commonly seen in cases of MCF in buffaloes. Furthermore, lymphocytic vasculitis centered in the adventitia, with occasional fibrinoid necrosis in the muscular layer, was found in the kidneys, liver, spleen, lymph nodes and brain. Nucleotide sequencing of DNA fragments from the central nervous system amplified by PCR revealed 98 percent similarity with known OHV-2 sequences from Genbank. Additionally, PCR analysis also revealed the presence of OHV-2 DNA in the peripheral mononuclear blood cells of two clinically healthy buffaloes. The diagnosis of MCFwas based on epidemiological, clinical, gross and histopathological findings and on the results of a semi-nested PCR followed by nucleotide sequencing.

É relatado um surto de febre catarral maligna (FCM) em Minas Gerais, que resultou na morte de 5 búfalas e uma vaca de uma mesma propriedade. Quatro búfalas morreram com 10-15 dias após o início dos sinais clínicos e uma búfala foi sacrificada in extremis, após manifestar sinais clínicos semelhantes. O exame histopatológico revelou lesões comumente observadas em búfalos com FCM como epicardite e miocardite histiolinfocítica multifocal e pneumonia linfocítica intersticial. Além disso, vasculite linfocítica, principalmente na camada adventícia, com necrose fibrinóide da camada muscular, foi observada no rim, fígado, baço, linfonodos e cérebro. A seqüência de nucleotídeos amplificada pela técnica de PCR revelou 98 por cento de homologia entre o fragmento de DNA amplificado da amostra do sistema nervoso central (SNC) da búfala com seqüências de OHV-2 previamente depositadas no Genbank. Adicionalmente, a técnica de PCR revelou a presença do DNA viral no sangue total de 2 búfalas, clinicamente sadias. O diagnóstico de FCM foi baseado em dados epidemiológicos, clínicos, patológicos, histopatológicos e semi-nested PCR.