Characterization of Caulobacter crescentus response to low temperature and identification of genes involved in freezing resistance

Free-living bacteria must respond to a wide range of temperature changes, and have developed specific mechanisms to survive in extreme environments. In this work we describe a remarkable resistance of mesophilic bacterium Caulobacter crescentus to several cycles of freezing at -80 degrees C, which was able to grow at low temperatures. Exponentially growing cells and late stationary-phase cells presented higher freezing resistance at both -20 and -80 degrees C than early stationary-phase cells. Cryotolerance was observed when log-phase cultures grown at 30 degrees C were preincubated at 5, 15 or 20 degrees C before freezing at -20 degrees C. A transposon library was screened to identify mutants sensitive to freezing at -80 degrees C and three strains presenting < 10% survival were isolated. Identification of genes disrupted in each mutant showed that they encoded an AddA family DNA helicase, a DEAD/DEAH box RNA helicase and a putative RND (resistance, nodulation, cell division) efflux system component. These strains showed longer generation times than wild-type cells when growing at 15 degrees C, with the RNA helicase mutant presenting a severe growth defect. These analyses suggest that the singular intrinsic resistance to freezing of C. crescentus is in fact a consequence of several independent traits, especially the maintenance of a proper degree of supercoiling of nucleic acids.

Resistance of melon cultivars to Bemisia tabaci biotype B

A mosca-branca, Bemisia tabaci biótipo B, é atualmente uma das mais importantes pragas da cultura do melão, ocasionando danos diretos e indiretos às plantas e reduzindo significativamente as produções a campo. Devido à necessidade de métodos mais sustentáveis do que o controle químico nas lavouras de melão, neste trabalho avaliou-se a resposta das cultivares de meloeiro AF-646, AF-682, Don Luis, Frevo, Jangada, Nilo, Vereda, Amarelo Ouro e Hales Best quanto à possível resistência a B. tabaci biótipo B, através de testes de campo, casa de vegetação e laboratório. No geral, 'Hales Best' e 'Amarelo Ouro' foram as mais resistentes, expressando não-preferência para oviposição contra a mosca-branca. A densidade de tricomas está associada à variação de oviposição sobre os materiais e deve ser melhor investigada em trabalhos futuros. Estes resultados podem auxiliar nos programas de melhoramento de melão, visando à resistência de plantas a B. tabaci biótipo B.

Resistance of stored bean varieties to Acanthoscelides obtectus (Coleoptera : Bruchidae)

During bean seed storage, yield can be lost due to infestations of Acanthoscelides obtectus Say, the bean weevil. The use of resistant varieties has shown promising results in fighting these insects, reducing infestation levels and eliminating chemical residues from the beans. The expression of resistance to A. obtectus in bean varieties is frequently attributed to the presence of phytohemagglutinins, protease inhibitors and alpha-amylase, and especially to variants of the protein arcelin, which reduce the larval viability of these insects. To evaluate the effect of bean seed storage time on the resistance expression of bean varieties to A. obtectus, tests with seeds of three ages (freshly-harvested, 4-month-old, and 8-month-old) were conducted in the laboratory, using four commercial varieties: Carioca Pitoco, Ipa 6, Porrillo 70, Onix; four improved varieties containing arcelin protein: Are. 1, Arc.2, Arc. 3, Arc.4; and three wild varieties also containing arcelin protein: Arc. IS, Arc.3S, and Arc. 5S. The Arc.5S, Arc. IS, and Arc.2 varieties expressed high antibiosis levels against the weevil; Arc. I and Arc3S expressed the same mechanism, but at lower levels. The occurrence of oviposition non-preference was also observed in Arc.5S and Arc. IS. The Arc.3 and Arc. 4 varieties expressed low feeding non-preference levels against A. obtectus. The expression of resistance in arcelin-bearing, wild or improved varieties was affected during the storage of seeds, and was high under some parameters but low in others. The results showed that addition of chemical resistance factors such as protein arcelin via genetic breeding may be beneficial in improving the performance of bean crops.

The field resistance of potato cultivars to foliar late blight and its relationship with foliage maturity type and skin type in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Anthelmintic resistance in cattle nematodes in northwestern São Paulo state, Brazil

The present study was carried out in the northwestern region of São Paulo State, Brazil, to determine the anthelmintic resistance status in cattle naturally infected with gastrointestinal nematodes. The anthelmintics tested were levamisole phosphate (Ripercol (R), Fort Dodge), albendazole sulphoxide (Ricobendazole, Fort Dodge), ivermectin (Ivomec (R), Merial) and moxidectin (Cydectin (R), Fort Dodge), administered at the doses recommended by the manufacturers. From April 2002 to May 2004, 25 cattle farms were evaluated. on each farm, steers were divided into treatment and control (not treated) groups based on fecal egg counts (FEC). Between 7 and 10 days after the anthelmintics administration, fecal samples were collected from each animal for post-treatment FEC. Fecal cultures from each group were also prepared for larval identification. After treatment, mean FEC reduction (FECR) in treatment groups (compared with control groups) was assessed on each farm. FECR was lower than 90% on 23 farms after ivermectin treatment. on 19 farms, FECR of 100% was recorded following moxidectin treatment; on the remaining 6, FECR ranged from 90% to 97.2%. After albendazole treatment, FECR was higher than 90% on 20 farms and ranged from 47.4% to 84.6% on other 5. After levamisole treatment, FECR was higher than 90% on 23 farms and equal to 47.4% and 73.7% on other 2 farms. Results indicated the presence of resistant Cooperia spp. and Haemonchus spp., especially to ivermectin; on some farms, resistance to albendazole and levamisole was also observed. (C) 2007 Elsevier B.V. All rights reserved.

Comparison of the susceptibility of Santa Ines and Ile de France ewes to nematode parasitism around parturition and during lactation

Breeds of sheep with resistance to nematode infection often display a reduced periparturient rise (PPR) in faecal egg counts (FEC) when compared with susceptible sheep. This trial was carried out to compare the resistance of Santa Ines, an indigenous Brazilian breed of sheep, and Ile de France ewes to natural infections by gastrointestinal nematodes, especially in the periparturient period. Twelve Santa lnes and 10 Ile de France ewes were estrus synchronized and then mated (December 2001). Lambing occurred from 30 April to 19 June 2002 and the lambs were weaned at 60 days of age. Faecal egg counts, packed cell volume (PCV), total plasma protein levels and peripheral eosinophils counts were determined from November 2001 to September 2002. To prevent deaths, individual treatment with anthelmintics was provided to ewes with FEC higher than 4000 eggs per gram (EPG) or with PCV lower than 21%. The percentage of Santa Ines ewes treated with anthelmintics was lower than that of Ile de France sheep. Eight of the 10 Ile de France ewes were treated, with one of them requiring four treatments, one requiring three treatments, and two requiring two treatments. The remaining four received a single treatment. In contrast, only 5 of the 12 Santa lnes ewes required treatment and for each this was done only once. The mean values of PCV were in the normal range in the Santa Ines ewes. In contrast, mean PCVs of Ile de France ewes reached a low of 24.5% in the second month after lambing and were significantly lower than Santa Ines 1 month before lambing (P < 0.05) and 2 months after lambing (P < 0.07). The mean plasma protein level was significantly lower in the Ile de France ewes in the second month after lambing (P < 0.01) coinciding with a reduction in PCV. The ewes of both breeds had high mean numbers of eosinophils in blood, which increased even further post-weaning when a marked reduction in FEC occurred. Haemonchus larvae were the most numerous genus in faecal cultures for both breeds followed by Trichostrongylus, Oesophagostomum and Cooperia. A periparturient rise in FEC was observed in both breeds. However, Santa Ines ewes showed a higher capacity to support the infection in an environment contaminated with large number of infective larvae of gastrointestinal nematodes. (C) 2004 Elsevier B.V. All rights reserved.

Evaluation of reduction in egg shedding of gastrointestinal nematodes in cattle following administration of anthelmintics

No intuito de avaliar o tempo necessário para redução dos valores de OPG visando identificar o início de atuação dos anti-helmínticos, cinco grupos de novilhos, naturalmente infectados por nematódeos gastrintestinais foram tratados com moxidectina, ivermectina, fosfato de levamisol e sulfóxido de albendazol. O levamisol promoveu redução no número de ovos de nematódeos eliminados nas fezes (R-OPG) de 97,4% 24 horas após a aplicação, a moxidectina de 98,3% após 36 horas, e o sulfóxido de albendazol de 95,9% após 36 horas. Foi registrada a presença de Cooperia spp. e Haemonchus spp. com resistência a ivermectina. A contagem de OPG realizada aos sete dias pós-tratamento apresentou resultados similares aos obtidos nas contagens realizadas 10 e 14 dias após a aplicação dos anti-helmínticos avaliados, demonstrando que o intervalo adequado entre o tratamento anti-helmíntico e o exame para verificar a redução do OPG pode ser de 7 dias.

Influence of protein supplementation during late pregnancy and lactation on the resistance of Santa Ines and Ile de France ewes to Haemonchus contortus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resistance of Santa Ines and crossbred ewes to naturally acquired gastrointestinal nematode infections

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

RESISTANCE OF TRYPANOSOMA-CRUZI TO BLOOD CLEARANCE INDUCED BY ACUTE-PHASE IMMUNE MOUSE SERUM

To investigate functional changes in Trypanosoma cruzi parasites induced during their interaction with the vertebrate host, we compared the blood clearance profiles of blood forms isolated from infected normal mice (Reg-Tc) or from infected mice immunodepressed after treatment with cyclophosphamide (Cy-Tc). Parasite blood numbers were measured at various time intervals in animals injected intravenously (i.v.) with 1-2 x 10(6) T. cruzi of either isolate. In the absence of added immune sera (spontaneous clearance), Reg-Tc and Cy-Tc were cleared from blood at similar rates. However, when acute immune mouse serum (Ac-IMS) was injected i.v. 2 min after inoculation of parasites, a significant proportion of Cy-Tc only was cleared from the blood an hour later, whereas Reg-Tc were not, their clearance profile being identical to that observed in mice injected with normal mouse serum. Cy-Tc susceptibility to Ac-IMS was not the result of a toxic effect of cyclophosphamide over T. cruzi as parasites recovered from animals immunodepressed by irradiation before infection were cleared similarly by acute serum. Contrary to Ac-IMS, chronic immune mouse serum induced similar rates of disappearance of Reg-Tc and Cy-Tc from blood. Our results suggest the occurrence of T. cruzi selection or modification during the acute phase, which leads to an increased parasite resistance to the clearance properties of acute-phase antibodies.

Staphylococcus aureus: Resistance patterns to antimicrobials and Penicillinase among strains isolated from apparently healthy lactating cows

Ninety-two strains of Staphylococcus aureus isolated from the nasal fossa and udder skin of apparently healthy lactating cows were analyzed for resistance to antibiotics and production of penicillinase.The results showed a greater frequency of resistance patterns to penicillin and ampicillin.All strains were sensitive to oxacyllin and gentamicin.The most frequent Barber and Burston model was SSSS (60.90%), followed by RSSS (18.50%).With respect to the production of penicillinase although the Lucas method indicated a larger number of positive samples, we suggest the use of the Haight and Finland method due to a greater consistency of data obtained with it.

Acquired resistance of horses to Amblyomma cajennense (Fabricius, 1787) ticks

Acquired immunity of horses to larvae, nymphs and adults of the Amblyomma cajennense tick was evaluated through three consecutive experimental infestations of tick-bite naive hosts. Data from these infestations were compared to those from field-sensitized horses and donkeys. It was observed that tick-bite naive horses developed a low level of resistance after two infestations as shown by a significant decrease in larval yield and a tendency for lower engorged weight of nymphs during third infestation. Ticks fed on field-sensitized horses had a similar biological performance to that observed on the third infestation of tick-bite naive horses but the mean engorged nymph weight was significantly lower than that of the first infestation from tick-bite naive horses. Donkeys presented the strongest resistance with significantly lower engorged weights of all instars and of the egg mass compared to the first infestation of tick-bite naive horses. Donkeys also displayed a significantly higher resistance than field-sensitized horses as demonstrated by significantly lower egg mass weights. Overall these results indicate that donkeys but not horses maintain a strong resistance to A. cajennense ticks. The importance of these findings in relation to vectoring of tick-borne diseases is discussed. (C) 2003 Elsevier B.V. All rights reserved.

Influence of the breed of bull (Bos taurus indicus vs. Bos taurus taurus) and the breed of cow (Bos taurus indicus, Bos taurus taurus and crossbred) on the resistance of bovine embryos to heat

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resistance of Santa Inês and Ile de France suckling lambs to gastrointestinal nematode infections.

A trial was carried out to determine the resistance to natural infection by gastrointestinal nematodes in 12 Santa Inês and nine Ile de France lambs before weaning. Faecal samples were obtained for faecal nematode egg counts (FEC). Blood samples were collected to determine packed cell volume (PCV), total plasma protein levels and peripheral eosinophil counts. Most Ile de France lambs (77.8%) were treated with an anthelmintic at 43 days of age, while 50% off Santa Inês lambs were treated at weaning, 57 days of age. The mean PCV values were normal in Santa Inês lambs, while in Ile de France lambs showed lower values reaching 22.3% at 43 days of age. The lowest mean plasma protein values were observed in Ile de France lambs (4.13 g/dl) at 43 days of age and in Santa Inês lambs (5.0 g/dl) at 57 days of age. Before weaning, Santa Inês lambs were susceptible to natural infections by gastrointestinal nematodes but with a greater capacity to stand the adverse effects of parasitism compared to Ile de France lambs.

Resistance of beef cattle of two genetic groups to ectoparasites and gastrointestinal nematodes in the state of São Paulo, Brazil

The resistance to infestations by ectoparasites and infections by gastrointestinal nematodes was studied in 45 animals (males and females) of two genetic groups: purebred Nelore (NI, n=28) and Three-Cross (1/2 Angus+1/4 Canchim+1/4 Nelore - TC, n=17). The animals were monitored for 24months, during which they were left to graze in tropical pastures without receiving treatment for parasites. Each month the animals were examined for infestations by external parasites, to count the numbers of cattle ticks Rhipicephalus microplus with diameter greater than 4.5mm present on the left side, horn flies (Haematobia irritans) present in the lumbar region and botfly larvae (Dermatobia hominis) present on the entire body. The H. irritans counts were performed with the aid of digital photographs. At the time of examination, fecal samples were collected to count the eggs per gram (EPG) and to perform coprocultures, and peripheral blood samples were drawn to determine the packed cell volume (PCV) and to count the eosinophils. For statistical analysis, the count data were transformed into log10 (n+1), where n is the number of parasites. For PCV, significant effects (P<0.05) were found for collection month (CO), genetic group (GG) and gender (SX), with means and respective standard errors of 41.5±0.65% for the NI animals, 39.3±0.83% for the TC, 41.5±0.72% for the females and 39.3±0.77% for the males. Regarding the eosinophil counts, only the effect of sex was significant (P<0.01), with means and respective standard errors of 926.0±46.2/μL, for males and 1088.0±43.8/μL of blood, for females. The NI animals presented lower mean counts for all the external parasites compared to the TC animals (P<0.01). For ticks, the transformed means followed by standard errors for the NI and TC animals were 0.06±0.01 and 0.34±0.02, while for horn flies these were 0.92±0.05 and 1.36±0.06 and for botfly larvae they were 0.05±0.03 and 0.45±0.05, respectively. The average EPG values were only influenced by CO (P<0.01). The coprocultures revealed the presence of the following endoparasites: Haemonchus spp., Cooperia spp., Oesophagostomum spp. and Trichostrongylus spp., the last in smaller proportion. There were no significant differences between the genetic groups for the endoparasite loads, except for Cooperia spp., which were present in greater number (P<0.05) in the NI group. The results obtained in this experiment confirm previous findings of greater susceptibility of the Nelore breed to Cooperia spp. and high resistance to ectoparasites. © 2013 Elsevier B.V.