Unlabelled: The objective of this field trial was to determine if vaccination against Haemophilus parasuis serovar 5 (HPS 5) and pathogenic serotypes of Escherichia coli would improve nursery pig performance in an outdoor unit in different seasons. The unit was concurrently infected with HPS 5 and with different serotypes of E. coli. All piglets were born to HPS 5 vaccinated sows. The trial was carried out in four (two summer and two winter) groups. Group 1 (E. coli and HPS vaccinated, summer season) (n = 362): Piglets were vaccinated pre-weaning with inactivated E. coli-VT2e-toxin and post-weaning against HPS 5. Group 2 (non-vaccinated, summer season) (n = 349): Piglets were not vaccinated. Group 3 (E. coli and HPS vaccinated, winter season) (n = 358): The animals were analogously treated as Group 1. Group 4 (non-vaccinated, winter season) (n = 353): Piglets were not vaccinated. The following parameters were evaluated: A: average daily nursery weight gain (ADG), B: nursery mortality, C: feed efficiency (FE). No significant weight differences were detected within the vaccinated and non-vaccinated summer or winter raised groups of weaners. Summer raised weaners were significantly (P<0.05) heavier from day 35 on than winter raised animals. ADG and FE of summer raised pigs were significantly better (weeks 1-3 P<0.05; fourth week post-weaning P<0.01) during the nursery period than that of the winter raised groups. Winter raised vaccinated weaners showed during the last week of nursing significantly (P<0.05) better daily gain and feed efficiency compared with the non-vaccinated winter raised animals. Non-significant ADG and FE differences were detectable between the summer raised vaccinated or non-vaccinated groups of pig. Winter raised non-vaccinated animals suffered significantly (P<0.05) higher nursery mortality (10.63%) compared to the winter raised vaccinated animals.
Implication: In cases of concurrent infections with HPS 5 and with different serotypes of E. coli, especially during winter season, vaccination against both diseases is suggested.
The studies were carried out on Balb/c mice (5-6 weeks of age) exposed to immunosuppression by a single intraperitoneal dose (125 mg/kg) of hydrocortisone. Prior to hydrocortisone injection the mice were treated with diethyldithiocarbamate (DTC) intra-peritoneally at a dose of 20 mg/kg, five times at 48 h intervals or calf thymus extract (TFX) at a dose of 10 mg/kg, 10 times at 24 h intervals. The two drugs were used per se or in zinc ions interactions, by adding zinc ions (as sulphate salt) to drinking water at a dose of 72 microg/mouse per day. The results obtained in the study show that hydrocortisone injection drastically decreases the number of thymocytes and splenocytes, which is also accompanied by a decreasing weight ratio of the thymus and spleen. The decreasing number of thymic and spleen cells corresponds to a decreasing percentage of CD4+, CD8+ and CD19+ splenocytes and double positive CD4+CD8+ thymocytes. Changes in the number of thymic cells affect their activity, which is expressed in a decreased proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohaemagglutinin (PHA). It has also been found that a single hydrocortisone dose decreases interleukin (IL)-1 production by murine intraperitoneal macrophages stimulated in vitro with lipopolysaccharide (LPS) from Escherichia coli. TFX or DTC counteract hydrocortisone-induced immunosuppression, which is expressed in partial normalization of the total number of thymic and spleen cells, accelerated regeneration of the two lymphatic organs, shorter suppressive action of hydrocortisone on the percentage of CD4+, CD8+ splenocytes and double positive (CD4+CD8+) and CD4+ thymocytes. Furthermore, total counteraction against the suppressive action of hydrocortisone to proliferative activity of thymocytes stimulated in vitro with Con A and PHA was observed. TFX administered prior to hydrocortisone injection partially prevented the suppressive action of the drug on IL-1 production by intraperitoneal macrophages, but such an effect was not observed with DTC. The immunorestorative effect of TFX and DTC was augmented by zinc supplementation. The results obtained in the study show that neither TFX nor DTC administration per se and in interaction with zinc supplementation were able to change the suppressive effect of hydrocortisone on the percentage of B splenocytes (CD19+ cells).
Twenty-two heifers were infected intranasally with non-cytopathic bovine viral diarrhoea virus (BVDV) between days 74 and 82 of pregnancy. All animals had developed serum antibodies against BVDV 5 weeks later. No clinical effects were seen in the heifers, and they all delivered a live calf. The newborn calves were generally small, appeared unthrifty as typical 'poor doers', and some developed secondary infections with diarrhoea and signs of respiratory disease. Eighteen of the 22 calves were born without antibodies against BVDV and were persistently infected (PI) with the virus. One was weak at birth and died the following day. Four calves were born with serum antibodies against BVDV and with no detectable virus. Three of these showed signs and/or pathological changes indicating disease in the central nervous system. Otherwise, there were no obvious clinical differences between these calves and the PI calves, nor were there any apparent significant differences in blood parameters between these groups. In general, the calves showed low gamma-globulin values and thrombocytopaenia, but moderately increased fibrinogen values and relatively normal lymphocyte numbers.
A culture technique employing cold enrichment at 4 degrees C followed by selective enrichment and plating at higher temperatures (30 degrees C) was used to isolate Listeria monocytogenes from faecal samples. The samples were held at 4 degrees C for 15 weeks and cultured weekly to assess the sensitivity of the culture after cold storage for different lengths of time. No media, Listeria selective enrichment broth (LSEB), nutrient broth (NB) and saline were used as cold storage medium. Cold storage increased the frequency of Listeria positive samples. The sensitivity of the culture for Listeria spp. and L. monocytogenes was 72 and 94%, and 56 and 61% after third and seventh week of cold storage, respectively. When the results of third and seventh week of cold storage were combined, the sensitivity was 100% for Listeria spp. and 94% for L. monocytogenes. LSEB and NB as storage medium increased Listeria positive samples after the first week of cold storage but did not maintain the increase thereafter while saline had an adverse effect on the growth of the bacteria. However, samples held in no media in a pilot study involving monthly sampling of a herd revealed better results. Detection limit of the culture media was also investigated. The lowest concentration detected by culture media was 3.17 organisms/ml. This was seven organisms/g for known Listeria positive sample. The faecal samples spiked with 10-fold dilutions of L. monocytogenes and held at 4 degrees C revealed that the sample spiked with 3.17 x 10-1 cfu/ml organisms resulted in growth after the second week of cold storage. The results suggest that the culture technique employing cold enrichment followed by selective enrichment and plating is more sensitive, the storage of faecal samples in no media when compared with the samples in storage medium, LSEB, NB and saline, during cold enrichment is a better application and culture of faeces, immediately after collection, at third and seventh week of cold enrichment produce more satisfactory results.
Mucosal disease (MD), one sequelae of bovine virus diarrhoea virus (BVDV) infection, causes severe lesions in lymphoid tissues and mucosal surfaces. Lesions are associated with the presence of cytopathogenic (cp) BVDV and initially characterized by apoptotic cell death. The objective of this investigation was to determine if this cell death is mediated only by the cp BVDV, which is known to induce apoptosis in cell culture or if immune-mediated host reactions might also contribute. Early onset MD was experimentally induced in calves by inoculation of persistently viremic calves with a closely related cp BVDV. Calves were euthanized in the early phase of infection between days 5 and 13 post-inoculation and tissues from tonsils, lymph nodes, Peyer's patches, jejunum and colon were collected. Presence of cp BVDV antigen was correlated with distribution of lymphocyte subpopulations in consecutive cryostat sections. In the lymphoid tissues, cp BVDV antigen was predominantly found in the lymphoid follicles. The increase of infected cells with time post-inoculation was paralleled by a decrease of B-lymphocytes and an increase of CD4+ T-lymphocytes. An increased number of CD8+ T-lymphocytes was seen in progressed lesions only. In the intestinal mucosa, initially multifocal, later diffuse infection with cp BVDV was accompanied by a multifocal or diffuse increase of CD4+ T-lymphocytes, respectively. Numbers of IgA+ plasma cells and CD8+ T-lymphocytes were decreased. The common change observed in lymphoid tissues and mucosa was the increase of CD4+ T-lymphocytes in sites with lesions. This might indicate a cell-mediated immune response to the cp BVDV. Besides their helper function to other cells of the immune system, activated CD4+ T-lymphocytes might also exert cytotoxic activity, induce apoptosis in target cells via Fas/Fas ligand binding and thus contribute to the severity of tissue lesions in MD.
The bactericidal effects of amoxicillin at below minimum inhibitory concentration (MIC) against Actinobacillus pleuropneumoniae NB001 were studied in vitro and in vivo. In vivo, the efficacy of amoxicillin on experimentally induced A. pleuropneumoniae infection in disease-free pigs was evaluated. Nine pigs were divided into three groups and all three groups were housed in the same room. Group I pigs were given long-acting amoxicillin injection 22 h prior to A. pleuropneumoniae challenge. Group II pigs were also A. pleuropneumoniae challenged but not given long-acting amoxicillin. Group III pigs were not treated. In vitro, A. pleuropneumoniae growth was suppressed in porcine blood with amoxicillin at below MIC. In vivo, clinical signs of disease were absent or mild in group I during 50 h post-challenge, and serum amoxicillin concentration was already less than MIC from 15 h post-challenge. Infected group II controls were severely affected by the infection, and mortality reached 100% within 50 h post-challenge. All non-treated pigs in group III became infected with NB001 from infected control pigs, and they displayed severe clinical signs of disease within 24 h post-challenge of groups I and II, and died within 50 h post-challenge of groups I and II.
Most human granulocytic ehrlichiosis (HGE) studies carried out in horses use needle inoculation of infected leucocytes or cell cultures. This route of inoculation does not accurately reflect natural infection of the tick-borne agent. To investigate whether tick transmission influences the course of granulocytic ehrlichiosis in the horse model, experimental transmission through infected laboratory-reared Ixodes scapularis ticks was attempted into two healthy horses. One additional horse served as negative control and was exposed to uninfected ticks. Eleven days after exposure to nymphal or adult ticks infected with Anaplasma phagocytophila (HGE agent) the two horses developed severe clinical and laboratory signs consistent with granulocytic ehrlichiosis. Bacteraemia was determined at various time points in the two horses by observation of morulae within neutrophils and by detection of A. phagocytophila genomic DNA by PCR of peripheral blood leucocytes. Further, both horses seroconverted. In contrast the control horse stayed uninfected. The results demonstrate that A. phagocytophila can be experimentally transmitted by infected nymphal and adult ticks and that the agent is able to produce a severe disease, similar to naturally occurring cases. Therefore, tick transmission is highly reproducible and can be successfully used in the equine animal model in order to study HGE.
Pharmacokinetic properties and tissue concentrations of enrofloxacin and ciprofloxacin were compared after intramuscular (i.m.) administrations of free and liposome-encapsulated enrofloxacin at the dose of 5 mg/kg body weight (bw). Twelve healthy adult New Zealand white rabbits were used in the experiment. Blood samples were obtained at 10, 20, 40, 60 and 90 min and 2, 4, 6, 8 and 12 h and tissue samples were collected 24 h after injection. Concentrations of drugs in serum were determined by high-performance liquid chromatography. Pharmacokinetics were best described by a two-compartment open model. Results indicated that absorption rate was slow, peak concentration was higher (P < 0.05), and the time to peak concentration (tmax congruent with 1.5 h) was significantly longer (P < 0.05) for liposome-encapsulated enrofloxacin (LEE) when compared with free enrofloxacin. Values of elimination half-life (t1/2beta = 12.9 h) and mean residence time (MRT = 17.6 h) of liposome-encapsulated enrofloxacin were longer (P < 0.05) and total clearance (Cl = 0.43 l/h/kg) was lower than those of free form. Moreover, the distribution volume at steady-state (Vd(ss) = 14.4 l/kg) of enrofloxacin administered encapsulated into liposomes was significantly higher (P < 0.05) than that of free enrofloxacin (FE). The tissue levels of enrofloxacin and ciprofloxacin after LEE injection were not different (P > 0.05) from FE. In conclusion, the result of present study suggest that LEE may be a beneficial and valuable formulation in the treatment of infectious diseases caused by sensitive pathogens in animals, providing sustained drug release from injection side and prolonged therapeutic serum concentrations after i.m. administration.
The German wasp, Vespula germanica has been observed to injure teats of dairy cows, causing lesions that are associated with clinical and subclinical mastitis. The presence of skin lesions on the teats, caused by the wasps, was recorded in a dairy cattle herd located in the Samaria foothills during July-October 1999. Wasp-inflicted injuries were found in 43.6% (58 of 133) of the adult cows and 1.4% (one of 71) of the first-calving cows. They were located in 42.4% of cows (n = 25) on all quarters, 18 cows (30.5%) had lesions on the front quarters and 27.1% (n = 16) of cows on the hind quarters only. Clinical and subclinical mastitis were diagnosed in 61% (36 of 59) and 28.8% (17 of 59), respectively, of the injured adult and first-calving cows. The most common bacterial isolates from the mastitic cows were Staphylococcus aureus 45.1% (n = 14), Streptococcus dysgalactiae 16.1% (n = 5), Streptococcus spp. 19.4% (n = 7) and others 13.9% (n = 5). The loss of milk production was estimated at 300 kg milk for each cow injured by wasps and exhibiting clinical mastitis. An increase in the bulk-milk somatic cell count, from 186 x 103 at 1 month prior to the outbreak to a peak of 1200 x 10(3) in the post-outbreak month, was noted. The culling rate reached 13.6% (eight of 59) of the affected cows. In summary, the considerable economic losses caused by the wasp infestation resulted from decreased milk production and a decline in milk quality, culling of affected cows, and increased demand for use of drugs and veterinary care.
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