Field evaluation of safety during gestation and horizontal spread of a recombinant differential bovine herpesvirus 1 (BoHV-1) vaccine

Bovine herpesvirus type 1 (BoHV-1) is recognized as a major cause of respiratory, reproductive disease and abortion in cattle. Vaccination is widely applied to minimize losses induced by BoHV-1 infections; however, vaccination of dams during pregnancy with modified live virus (MLV) vaccines has been occasionally associated to abortions. We have previously reported the development of a BoHV-1 recombinant virus, constructed with basis on a Brazilian BoHV-1 (Franco et al. 2002a) from which the gene coding for glycoprotein E (gE) was deleted (gE-) by genetic manipulation. Such recombinant has been previously evaluated in its potential as a differential vaccine (gE- vaccine) that allows differentiation between vaccinated and infected animals. Here, in the first part of the present study, the safety of the gE- vaccine during pregnancy was evaluated by the intramuscular inoculation of 10(7.4) tissue culture 50 % infective doses (TCID50) of the virus into 22 pregnant dams (14 BoHV-1 seronegative; 8 seropositive), at different stages of gestation. Other 15 pregnant dams were kept as non-vaccinated controls. No abortions, stillbirths or fetal abnormalities were seen after vaccination. Seroconversion was observed in both groups of previously seronegative vaccinated animals. In the second part of the study, the potential of the gE- vaccine virus to spread among beef cattle under field conditions was examined. Four heifers were inoculated intranasally with a larger amount (10(7,6) TCID50) of the gE- vaccine (to increase chances of transmission) and mixed with other sixteen animals at the same age and body condition, in the same grazing area, at a population density equal to the average cattle farming density within the region (one cattle head per 10,000 m²), for 180 days. All animals were monitored daily for clinical signs. Serum samples were collected on days 0, 30, 60 and 180 post-vaccination. Seroconversion was observed only in vaccinated heifers. These results indicate that, under the conditions of the present study, the gE- vaccine virus did not cause any noticeable harmful effect on pregnant dams and on its offspring and did not spread horizontally among cattle.

Importância do eletrorretinograma de campo total (Full field ERG) em cães da raça Cocker Spaniel Inglês portadores de catarata

Com o aumento do número de intervenções cirúrgicas para a remoção da catarata em cães, observa-se a necessidade de exames específicos que devem ser realizados antes da indicação cirúrgica. A catarata é uma das principais causas de cegueira em cães. Nos estágios mais avançados, impossibilita o exame fundoscópico e inviabiliza a observação de alterações retinianas como a atrofia progressiva da retina (APR), degeneração retiniana hereditária de acometimento bilateral que, quando diagnosticada, contra-indica a cirurgia. Com o intuito de reestabelecer a visão, o eletrorretinograma de campo total (Full field ERG) torna-se indispensável na avaliação pré-cirúrgica da remoção da catarata. Como os cães da raça Cocker Spaniel Inglês são predispostos à catarata e a degenerações retinianas, objetivamos neste estudo avaliar as respostas dos ERGs realizados nestes animais. Foram avaliados 136 eletrorretinogramas de cães da raça Cocker Spaniel Inglês (62 machos e 74 fêmeas, com idades entre 3 e 15 anos) no período de Setembro de 2004 a Maio de 2009. Todos os animais apresentavam baixa de visão e catarata durante o exame. O diagnóstico de degeneração retiniana foi baseado nos valores de amplitude pico a pico e tempo de culminação da onda-b nas 3 respostas (resposta escotópica de bastonetes, máxima resposta e resposta fotópica de cones), idade do paciente e estágio de maturação da catarata. Nos cães sem degeneração retiniana, a média da amplitude e do tempo de culminação nas três respostas obtidas foram, respectivamente: 71,55mV/65,15ms; 149,17mV/33,03ms; 31,06mV/27,90ms. Nos cães com degeneração retiniana, 38 animais apresentaram ERG extinto. Dentre os restantes que apresentavam baixas respostas, a média da amplitude e do tempo de culminação nas três respostas obtidas foram, respectivamente: 12,88mV/65,04ms; 24,16mV/36,25ms; 8,36mV/31,38ms. Foi observado que em 122 animais (89,7%) os exames eram compatíveis com diagnóstico de atrofia progressiva da retina. Frente aos resultados obtidos, conclui-se que os cães da raça Cocker Spaniel Inglês portadores de catarata devem ser submetidos ao eletrorretinograma de campo total antes da remoção cirúrgica devido à alta incidência de degeneração retiniana nesta raça.

Comparison of full-field electroretinogram in diabetic and non diabetic dogs with cataracts

Being the commonest ocular disorder, dense cataracts disable fundoscopic examination and the diagnosis of retinal disorders, which dogs may be predisposed. The aim of this study was to compare the electroretinographic responses recorded according to the International Society for Clinical Electrophysiology of Vision human protocol to evaluate retinal function of diabetic and non diabetic dogs, both presenting mature or hypermature cataracts. Full-field electroretinogram was recorded from 66 dogs, with ages varying from 6 to 15 years old allocated into two groups: (1) CG, non diabetic cataractous dogs, and (2) DG, diabetic cataractous dogs. Mean peak-to-peak amplitude (microvolts) and b-wave implicit time (milliseconds) were determined for each of the five standard full-field ERG responses (rod response, maximal response, oscillatory potentials, single-flash cone response and 30 Hz flicker). Comparing CG to DG, ERGs recorded from diabetic dogs presented lower amplitude and prolonged b-wave implicit time in all ERG responses. Prolonged b-wave implicit time was statistically significant (p< 0.05) at 30 Hz flicker (24.0 ms versus 22.4 ms). These data suggests full-field ERG is capable to record sensible alterations, such as flicker's implicit time, being useful to investigate retinal dysfunction in diabetic dogs.

Weak phenotypic reversion of ivermectin resistance in a field resistant isolate of Haemonchus contortus by verapamil

Recent advances in anthelmintic resistant phenotype reversion by Pgp modulating drugs in ruminant nematodes indicate that this can be a useful tool to helminth control. The aim of the present study was to evaluate the efficacy of ivermectin (IVM) in combination with verapamil (VRP), in oil or water-based vehicle, against an IVM-resistant field isolate of Haemonchus contortus through a larval migration assay and experimental infection trial. In the in vitro assay was observed a phenotypic reversion of H. contortus resistance to ivermectin at a high concentration of VRP, increasing IVM efficacy from 53.1% to 94.3. In the in vivo trial, IVM + VRP demonstrated 36.02% efficacy compared to the 7.75% of IVM alone. The vehicle formulation showed no influence in efficacy. These are the first results demonstrating the effect of VRP as a partial IVM-resistance phenotype reverser in a field isolate of IVM-resistant H. contortus experimentally inoculated in sheep.

Cysteine addition on short-term cooled boar semen preservation and its relationship with swine field fertility

Artificial insemination is routinely used in the swine industry to reduce the costs of production through to increase the efficiency of the refrigerated boar semen process. The objective of this study was to evaluate the effect of different levels of cysteine (CYS) added to the Beltsville Thawing Solution (BTS) extender semen during cooling for up to 72 hours. Ejaculated from three boars were collected with the gloved-hand technique and semen aliquots were diluted in BTS as follow: BTS only (BTS), BTS + 0.1mM cysteine (CYS0.1), BTS + 0.5mM cysteine (CYS0.5), BTS + 1.0mM cysteine (CYS1.0), BTS + 2.5mM cysteine (CYS2.5), BTS + 5.0mM cysteine (CYS5.0), BTS + 10.0mM cysteine (CYS10.0), and BTS + 20.0mM cysteine (CYS20.0). Evaluation of sperm integrity were analyzed using 0.5mg/ml propidium iodide (plasma membrane), 100µg/ml isothiocynate-conjugated Pisum sativun agglutinin (acrosomal membrane) and 153µM 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanine iodide (mitochondria potential) after semen dilution at specific times (0, 24, 48 and 72 hours). Additionally, we also evaluated the effects of 5.0 mM CYS addition in the BTS extender on the maintenance of sperm quality and their influence on fertility in the swine production. After artificial insemination, animals were evaluated based on the estrous return and the number of piglet's born. Cysteine at concentrations of 10.0 and 20.0mM resulted in more pronounced reductions even at the time zero. Semen viability decreased to levels below 10% at these high levels of CYS in the first 24 hour of storage at 17ºC. At the end of the storage time, less than 65% of sperm cells had intact plasma membrane in all groups. The sperm viability decreased significantly when the semen was added at high concentrations of CYS (time "0"; CYS10.0 and CYS20.0; p<0.05), when compared to the other CYS concentrations. The BTS (10.20±0.39) treated group showed a lower rate of estrus return when compared to other (BTSCYS; 86.05±039), and it showed also the highest total number of piglets borne per treatment (12.71±3.38 vs. 9.00±3.38, respectively). In conclusion, the addition of CYS in the BTS semen extender did not maintain spermatic viability of boar cooled spermatozoa and it results in a higher percentage of return to estrus and lower number of piglets borne.

Evaluation of resistance in a selected field strain of Haemonchus contortus to ivermectin and moxidectin using the Larval Migration on Agar Test

Haemonchus contortus is one of the most common and economically significant causes of disease in small ruminants worldwide, and the control programs of parasitic nematodes - including H. contortus - rely mostly on the use of anthelmintic drugs. The consequence of the use of this, as the sole sanitary strategy to avoid parasite infections, was the reduction of the efficacy of all chemotherapeutic products with a heavy selection for resistance. The widespread of anthelmintic resistance and the difficulty of its early diagnosis has been a major concern for the sustainable parasite management on farms. The objective of this research was to determine and compare the ivermectin (IVM) and moxidectin (MOX) effect in a selected field strain of H. contortus with a known resistance status, using the in vitro larval migration on agar test (LMAT). Third stage larvae of the selected isolate were obtained from faecal cultures of experimentally infected sheep and incubated in eleven increasing diluted concentrations of IVM and MOX (6, 12, 24, 48, 96, 192, 384, 768, 1536, 3072 and 6144µg/mL). The dose-response sigmoidal curves were obtained using the R² value of >0.90 and the lethal concentration (LC50) dose for the tested anthelmintic drugs using a four-parameter logistic model. The LC50 value for MOX was significantly lower than IVM (1.253µg/mL and 91.06µg/mL), identifying the H. contortus isolate as considerably less susceptible to IVM compared to MOX. Furthermore, the LMAT showed a high consistency (p<0.0001) and provided to be a useful diagnostic tool for monitoring the resistance status of IVM and MOX in H. contortus field isolate, as well as it may be used for official routine drug monitoring programs under the Ministry of Agriculture (MAPA) guidance.

Bus bar aeolian vibration field tests

Large amplitude bus bar aeolian vibration may lead to post insulator damage. Different damping applications are used to decrease the risk of large amplitude aeolian vibration. In this paper the post insulator load caused by the bus bar aeolian vibration and the effect of damping methods are evaluated. The effects of three types of bus bar connectors and three types of primary structures are studied. Two actual damping devices, damping cable and their combinations are studied. The post insulator loads are studied with strain gage based custom made force sensors installed on the both ends of the post insulator and with the displacement sensor installed on the midpoint of the bus bar. The post insulator loads are calculated from the strain values and the damping properties are determined from the displacement history. The bus bar is deflected with a hanging weight. The weight is released and the bus bar is left to free damped vibration. Both actual bus bar vibration dampers RIBE and SBI were very effective against the aeolian vibration. Combining vibration damper with damping cable will increase the damping ratio but it may be unnecessary considering the extra effort. Bus bar connector type or primary structure have no effect on the vertical load. The bending moment at the post insulator with double sided bus bar connector is significantly higher than at the post insulator with single sided bus bar connector. No reliable conclusions about bus bar connector type effect can be done, but the roller bearing type or central bearing type connector may reduce the bending moment. The RHS steel frame as primary structure may increase the bending moment peak values since it is the least rigid primary structure type and it may start to vibrate as a response to the awakening force of the vibrating bus bar.