Objective: To determine the reference interval for WBC counts in Holstein dairy cows from herds with high seroprevalence for anti–bovine leukemia virus (BLV) antibodies, analyze the correlation of total WBC counts and blood proviral load (bPVL) in BLV-infected animals, and determine whether total WBC count can be used a hematologic marker for in vivo infection. Animals: 307 lactating cows from 16 dairy herds with high BLV seroprevalence. Procedures: Blood samples were collected for assessment of plasma anti–BLV p24 antibody concentration (all cows), manual determination of WBC count (161 BLV-seronegative cows from 15 herds), and evaluation of bPVL (146 cows from another herd). Results: The WBC count reference interval (ie, mean ± 2 SD) for BLV-seronegative dairy cows was 2,153 to 11,493 cells/μL. Of the 146 cows used to analyze the correlation between WBC count and bPVL, 107 (73%) had WBC counts within the reference interval; of those cows, only 21 (19.6%) had high bPVL. Most cows with high WBC counts (35/39) had high bPVL. Mean WBC count for cows with high bPVL was significantly higher than values for cows with low or undetectable bPVL. White blood cell counts and bPVL were significantly (ρ = 0.71) correlated. Conclusions and Clinical Relevance: These data have provided an updated reference interval for WBC counts in Holstein cows from herds with high BLV seroprevalence. In dairy cattle under natural conditions, WBC count was correlated with bPVL; thus, WBC count determination could be a potential tool for monitoring BLV infection levels in attempts to control transmission.

Objective—To determine the reference interval for WBC counts in Holstein dairy cows from herds with high seroprevalence for anti–bovine leukemia virus (BLV) antibodies, analyze the correlation of total WBC counts and blood proviral load (bPVL) in BLV-infected animals, and determine whether total WBC count can be used a hematologic marker for in vivo infection. Animals—307 lactating cows from 16 dairy herds with high BLV seroprevalence. Procedures—Blood samples were collected for assessment of plasma anti–BLV p24 antibody concentration (all cows), manual determination of WBC count (161 BLV-seronegative cows from 15 herds), and evaluation of bPVL (146 cows from another herd). Results—The WBC count reference interval (ie, mean ± 2 SD) for BLV-seronegative dairy cows was 2,153 to 11,493 cells/μL. Of the 146 cows used to analyze the correlation between WBC count and bPVL, 107 (73%) had WBC counts within the reference interval; of those cows, only 21 (19.6%) had high bPVL. Most cows with high WBC counts (35/39) had high bPVL. Mean WBC count for cows with high bPVL was significantly higher than values for cows with low or undetectable bPVL. White blood cell counts and bPVL were significantly (ρ = 0.71) correlated. Conclusions and Clinical Relevance—These data have provided an updated reference interval for WBC counts in Holstein cows from herds with high BLV seroprevalence. In dairy cattle under natural conditions, WBC count was correlated with bPVL; thus, WBC count determination could be a potential tool for monitoring BLV infection levels in attempts to control transmission. | Instituto de Virología | Fil: Alvarez, Irene. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Gutierrez, Gerónimo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Gammella, Mariela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Martinez, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Politzki, Romina Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Gonzalez, Cintia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Caviglia, Luciana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Carignano, Hugo Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina | Fil: Fondevila, Norberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina | Fil: Poli, Mario Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética "Ewald A. Favret"; Argentina | Fil: Trono, Karina Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina

Objective: To measure florfenicol concentrations in ovine tear fluid after IM and SC administration and determine minimum inhibitory concentrations (MICs) of florfenicol against field isolates of Mycoplasma organisms potentially involved in infectious keratoconjunctivitis. Animals: 9 healthy adult Lacaune ewes. Procedures: Animals received an IM and SC administration of florfenicol (20 mg/kg) in a 2-way crossover design. Samples of blood and tear fluid were collected before and for 24 hours after administration. Concentrations of florfenicol in plasma and tear fluid were measured via high-performance liquid chromatography. The MIC of florfenicol for various Mycoplasma strains cultured from sheep and goats was determined via an agar dilution method. Results: Mean florfenicol concentration in tear fluid for the 24-hour period was significantly higher after IM administration (0.70 μg/mL) than after SC administration (0.22 μg/mL) and was maintained for a longer duration. The lacrimal fluid-to-plasma concentration ratio was not different between the 2 routes of administration, with mean values of 40.2% and 32.5% after IM and SC administration, respectively. The MIC for Mycoplasma agalactiae, Mycoplasma conjunctivae, and Mycoplasma mycoides isolates ranged from 0.5 to 8 μg of florfenicol/mL. Two strains of M agalactiae could be considered resistant to florfenicol. Conclusions and Clinical Relevance: Florfenicol readily penetrated the preocular tear fluid of sheep after IM and SC administration. For both routes of administration, doses > 20 mg/kg would be necessary to achieve tear fluid concentrations of florfenicol greater than the MICs for most strains of Mycoplasma organisms.

To measure florfenicol concentrations in ovine tear fluid after IM and SC administration and determine minimum inhibitory concentrations (MICs) of florfenicol against field isolates of Mycoplasma organisms potentially involved in infectious keratoconjunctivitis. [br/]ANIMALS: 9 healthy adult Lacaune ewes. [br/]PROCEDURES: Animals received an IM and SC administration of florfenicol (20 mg/kg) in a 2-way crossover design. Samples of blood and tear fluid were collected before and for 24 hours after administration. Concentrations of florfenicol in plasma and tear fluid were measured via high-performance liquid chromatography. The MIC of florfenicol for various Mycoplasma strains cultured from sheep and goats was determined via an agar dilution method. [br/]RESULTS: Mean florfenicol concentration in tear fluid for the 24-hour period was significantly higher after IM administration (0.70 μg/mL) than after SC administration (0.22 μg/mL) and was maintained for a longer duration. The lacrimal fluid-to-plasma concentration ratio was not different between the 2 routes of administration, with mean values of 40.2% and 32.5% after IM and SC administration, respectively. The MIC for Mycoplasma agalactiae, Mycoplasma conjunctivae, and Mycoplasma mycoides isolates ranged from 0.5 to 8 μg of florfenicol/mL. Two strains of M agalactiae could be considered resistant to florfenicol. [br/]CONCLUSIONS AND CLINICAL RELEVANCE: Florfenicol readily penetrated the preocular tear fluid of sheep after IM and SC administration. For both routes of administration, doses > 20 mg/kg would be necessary to achieve tear fluid concentrations of florfenicol greater than the MICs for most strains of Mycoplasma organisms.

To measure florfenicol concentrations in ovine tear fluid after IM and SC administration and determine minimum inhibitory concentrations (MICs) of florfenicol against field isolates of Mycoplasma organisms potentially involved in infectious keratoconjunctivitis. [br/]ANIMALS: 9 healthy adult Lacaune ewes. [br/]PROCEDURES: Animals received an IM and SC administration of florfenicol (20 mg/kg) in a 2-way crossover design. Samples of blood and tear fluid were collected before and for 24 hours after administration. Concentrations of florfenicol in plasma and tear fluid were measured via high-performance liquid chromatography. The MIC of florfenicol for various Mycoplasma strains cultured from sheep and goats was determined via an agar dilution method. [br/]RESULTS: Mean florfenicol concentration in tear fluid for the 24-hour period was significantly higher after IM administration (0.70 μg/mL) than after SC administration (0.22 μg/mL) and was maintained for a longer duration. The lacrimal fluid-to-plasma concentration ratio was not different between the 2 routes of administration, with mean values of 40.2% and 32.5% after IM and SC administration, respectively. The MIC for Mycoplasma agalactiae, Mycoplasma conjunctivae, and Mycoplasma mycoides isolates ranged from 0.5 to 8 μg of florfenicol/mL. Two strains of M agalactiae could be considered resistant to florfenicol. [br/]CONCLUSIONS AND CLINICAL RELEVANCE: Florfenicol readily penetrated the preocular tear fluid of sheep after IM and SC administration. For both routes of administration, doses > 20 mg/kg would be necessary to achieve tear fluid concentrations of florfenicol greater than the MICs for most strains of Mycoplasma organisms.

A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n= 154) from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822) were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74%(89/121) were infected with trypanosomes from the Vaal Dam and 63%(63/100) from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25%(1/4) was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2) prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa) and the Sand River Dam (Swaziland). Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record.

A total of 257 fishes from four families, Clariidae, Cichlidae, Cyprinidae and Schilbeidae were collected from three localities: the Sand River Dam, Swaziland; the Nylsvlei Nature Reserve, South Africa and the Vaal Dam and Vaal River Barrage, South Africa. Only fishes (n= 154) from Clariidae and Cichlidae were found to be infected with trypanosomes. A total of 221 Clarias gariepinus (Burchell 1822) were collected from the Vaal Dam and Vaal Barrage area, South Africa. Of these, 74%(89/121) were infected with trypanosomes from the Vaal Dam and 63%(63/100) from the Vaal River Barrage, with no seasonal infection pattern. A prevalence of 25%(1/4) was found in C. gariepinus from the Sand River Dam, Swaziland, and a 50% (1/2) prevalence was found in Tilapia sparrmanii from the Nylsvlei Nature Reserve, South Africa. Standard measurements conformed closely to the morphometric and morphological descriptions of Trypanosoma mukasai. This article provides new locality records for T. mukasai from the Vaal Dam, Vaal River Barrage and Nylsvlei Nature Reserve (South Africa) and the Sand River Dam (Swaziland). Tilapia sparrmanii collected in the Sand River Dam in Swaziland is also noted as a new host record.

A retrospective serosurvey was carried out between 2009 and 2012 to detect antibodies to Brucella spp. in free-ranging African wildlife ungulates from five selected game parks in Zimbabwe. Samples were drawn from wildlife-livestock interface and non-interface areas in Zimbabwe. A total of 270 serum samples from four different species, namely African buffalo (Syncerus caffer) (n=106), impala (Aepyceros melampus) (n = 72), black rhinoceros (Diceros bicornis) (n= 45) and white rhinoceros (Ceratotherium simum) (n = 47), were tested. The percentage of positive samples was 17.0% in buffalo (18/106; 95% CI: 9.72% – 24.1%) and 1.4% in impala (1/72; 95% CI: 0% – 4.2%). No antibodies to Brucella spp. were detected in the two rhinoceros species. The difference in the percentage of seropositive cases between buffalo and impala was significant (p< 0.05). Seropositivity to Brucella spp. was higher (19.1%) in adult buffalo compared with juveniles and sub-adults younger than six years (5.9%). Further, seropositivity was marginally higher (20.4%) in animals from wildlife-livestock interface areas than in those from non-interface areas (13.45%; OR = 1.45) although the difference was not statistically significant. The study showed that brucellosis could be more widespread in buffalo and may circulate in this species independently in the absence of contact with cattle, whilst rhinoceros may be considered less susceptible to brucellosis. The role of the wildlife-livestock interface in the epidemiology of brucellosis in wildlife and livestock is probably overstated but needs to be explored further.

A retrospective serosurvey was carried out between 2009 and 2012 to detect antibodies to Brucella spp. in free-ranging African wildlife ungulates from five selected game parks in Zimbabwe. Samples were drawn from wildlife-livestock interface and non-interface areas in Zimbabwe. A total of 270 serum samples from four different species, namely African buffalo (Syncerus caffer) (n=106), impala (Aepyceros melampus) (n = 72), black rhinoceros (Diceros bicornis) (n= 45) and white rhinoceros (Ceratotherium simum) (n = 47), were tested. The percentage of positive samples was 17.0% in buffalo (18/106; 95% CI: 9.72% – 24.1%) and 1.4% in impala (1/72; 95% CI: 0% – 4.2%). No antibodies to Brucella spp. were detected in the two rhinoceros species. The difference in the percentage of seropositive cases between buffalo and impala was significant (p 0.05). Seropositivity to Brucella spp. was higher (19.1%) in adult buffalo compared with juveniles and sub-adults younger than six years (5.9%). Further, seropositivity was marginally higher (20.4%) in animals from wildlife-livestock interface areas than in those from non-interface areas (13.45%; OR = 1.45) although the difference was not statistically significant. The study showed that brucellosis could be more widespread in buffalo and may circulate in this species independently in the absence of contact with cattle, whilst rhinoceros may be considered less susceptible to brucellosis. The role of the wildlife-livestock interface in the epidemiology of brucellosis in wildlife and livestock is probably overstated but needs to be explored further.

Several outbreaks of Rift Valley fever (RVF) have been documented in South Africa since it first occurred in the country in 1950. However, there is no comprehensive account of the timing, location and extent of all known outbreaks. As part of a study investigating the epidemiology of RVF in South Africa, a full history of outbreaks was compiled using references to the disease in South Africa from scientific literature, annual reports, disease reports and animal disease databases. The geographic location and temporal occurrence of each outbreak were recorded as accurately as allowed by the available records. The result was a better and more complete picture than has hitherto been available of the spatial and temporal distribution of RVF in South Africa for the period between 1950 and 2011. Several smaller outbreaks which had not been described previously in literature were documented. Extensive outbreaks occurred in the central interior of the country (Free State, Eastern Cape and Northern Cape provinces), interspersed with smaller outbreaks or long intervening periods of absence, whilst smaller outbreaks occurred in the eastern part of the country (KwaZulu-Natal, Mpumalanga and Gauteng).

Several outbreaks of Rift Valley fever (RVF) have been documented in South Africa since it first occurred in the country in 1950. However, there is no comprehensive account of the timing, location and extent of all known outbreaks. As part of a study investigating the epidemiology of RVF in South Africa, a full history of outbreaks was compiled using references to the disease in South Africa from scientific literature, annual reports, disease reports and animal disease databases. The geographic location and temporal occurrence of each outbreak were recorded as accurately as allowed by the available records. The result was a better and more complete picture than has hitherto been available of the spatial and temporal distribution of RVF in South Africa for the period between 1950 and 2011. Several smaller outbreaks which had not been described previously in literature were documented. Extensive outbreaks occurred in the central interior of the country (Free State, Eastern Cape and Northern Cape provinces), interspersed with smaller outbreaks or long intervening periods of absence, whilst smaller outbreaks occurred in the eastern part of the country (KwaZulu-Natal, Mpumalanga and Gauteng).

The attenuated <em>Salmonella typhimurium χ</em>4550 strain was used to harbour a reconstructed bicistronic DNA vaccine against porcine rotavirus, which carried the rotavirus nonstructural protein 4 (<em>NSP4</em>) and VP7 genes simultaneously. Using a balanced lethal system, the kanamycin resistance gene of expressing eukaryotic plasmids pVAX1 and pVAXD were replaced by the aspartate β-semialdehyde dehydrogenase (<em>asd</em>) gene. The NSP4 cleavage product (259–525) of rotavirus OSU strain and VP7 full-length genes were amplified by reverse transcription polymerase chain reaction and then inserted into the eukaryotic single-expression plasmid, pVAX1-asd, and the eukaryotic dual-expression plasmid, pVAXD-asd, respectively. The recombinant plasmids pVAX1-asd-NSP4, pVAX1-asd-VP7 and pVAXD-asd-NSP4-VP7 were transformed into the attenuated <em>S. typhimurium χ</em>4550 strain by electrotransformation. An indirect immunofluorescence assay of the expressed COS-7 cell suggested that the recombinant <em>S. typhimurium χ</em>4550 strain was constructed successfully. The recombinant <em>S. typhimurium χ</em>4550 strain was orally administered to BALB/c mice. The group immunised with dual- expression plasmids produced a significantly higher level of serum Immunoglobulin G (IgG) and intestinal Immunoglobulin A (IgA) than the group immunised with single-expression plasmids. These results indicated that eukaryotic bicistronic plasmid DNA vaccines could be successfully constructed to enhance humoural, mucosal and cellular immune response against rotavirus infection.

The attenuated Salmonella typhimurium χ4550 strain was used to harbour a reconstructed bicistronic DNA vaccine against porcine rotavirus, which carried the rotavirus nonstructural protein 4 (NSP4) and VP7 genes simultaneously. Using a balanced lethal system, the kanamycin resistance gene of expressing eukaryotic plasmids pVAX1 and pVAXD were replaced by the aspartate β-semialdehyde dehydrogenase (asd) gene. The NSP4 cleavage product (259–525) of rotavirus OSU strain and VP7 full-length genes were amplified by reverse transcription polymerase chain reaction and then inserted into the eukaryotic single-expression plasmid, pVAX1-asd, and the eukaryotic dual-expression plasmid, pVAXD-asd, respectively. The recombinant plasmids pVAX1-asd-NSP4, pVAX1-asd-VP7 and pVAXD-asd-NSP4-VP7 were transformed into the attenuated S. typhimurium χ4550 strain by electrotransformation. An indirect immunofluorescence assay of the expressed COS-7 cell suggested that the recombinant S. typhimurium χ4550 strain was constructed successfully. The recombinant S. typhimurium χ4550 strain was orally administered to BALB/c mice. The group immunised with dual- expression plasmids produced a significantly higher level of serum Immunoglobulin G (IgG) and intestinal Immunoglobulin A (IgA) than the group immunised with single-expression plasmids. These results indicated that eukaryotic bicistronic plasmid DNA vaccines could be successfully constructed to enhance humoural, mucosal and cellular immune response against rotavirus infection.

In order to get a reliable estimate of brucellosis prevalence in Eritrean dairy cattle, a cross-sectional study was carried out in 2009. The survey considered the sub-population of dairy cattle reared in modern small- and medium-sized farms. Samples were screened with the Rose Bengal test (RBT) and positive cases were confirmed with the complement fixation test (CFT). A total of 2.77%(417/15 049; Credibility Interval CI: 2.52% – 3.05%) of the animals tested in this study were positive for antibodies to Brucellaspecies, with a variable and generally low distribution of positive animals at regional level. The highest seroprevalence was found in the Maekel region (5.15%; CI: 4.58% – 5.80%), followed by the Debub (1.99%; CI: 1.59% – 2.50%) and Gash-Barka (1.71%; CI: 1.34% – 2.20%) regions. Seroprevalence at sub-regional levels was also generally low, except for two sub-regions of Debub and the sub-region Haicota from the Gash-Barka region. Seroprevalence was high and more uniformly distributed in the Maekel region, namely in the Asmara, Berik and Serejeka sub-regions. Considering the overall low brucellosis prevalence in the country, as identified by the present study, a brucellosis eradication programme for dairy farms using a test-and-slaughter policy would be possible. However, to encourage the voluntary participation of farmers to the programme and to raise their awareness of the risks related to the disease for animals and humans, an extensive public awareness campaign should be carefully considered, as well as strict and mandatory dairy movement control.

In order to get a reliable estimate of brucellosis prevalence in Eritrean dairy cattle, a cross-sectional study was carried out in 2009. The survey considered the sub-population of dairy cattle reared in modern small- and medium-sized farms. Samples were screened with the Rose Bengal test (RBT) and positive cases were confirmed with the complement fixation test (CFT). A total of 2.77%(417/15 049; Credibility Interval CI: 2.52% – 3.05%) of the animals tested in this study were positive for antibodies to Brucellaspecies, with a variable and generally low distribution of positive animals at regional level. The highest seroprevalence was found in the Maekel region (5.15%; CI: 4.58% – 5.80%), followed by the Debub (1.99%; CI: 1.59% – 2.50%) and Gash-Barka (1.71%; CI: 1.34% – 2.20%) regions. Seroprevalence at sub-regional levels was also generally low, except for two sub-regions of Debub and the sub-region Haicota from the Gash-Barka region. Seroprevalence was high and more uniformly distributed in the Maekel region, namely in the Asmara, Berik and Serejeka sub-regions. Considering the overall low brucellosis prevalence in the country, as identified by the present study, a brucellosis eradication programme for dairy farms using a test-and-slaughter policy would be possible. However, to encourage the voluntary participation of farmers to the programme and to raise their awareness of the risks related to the disease for animals and humans, an extensive public awareness campaign should be carefully considered, as well as strict and mandatory dairy movement control.

<span>In 2004, a new concept was introduced for simplifying identification of larvae of the common nematodes of cattle, sheep and goats that comprises estimates of the lengths of the sheath tail extensions of infective third-stage larvae (L</span>₃<span>) of each genus and/or species to that of </span><em>Trichostrongylus</em><span> spp., instead of having to be dependent only on measurements in micrometre. For example, if the mean length of the sheath tail extension (the extension of the sheath caudad, beyond the caudal tip of the larva) of </span><em>Trichostrongylus colubriformis </em><span>and</span><em> Trichostrongylus axei</em><span> is assumed to be ‘X’, then that of</span><em>Haemonchus contortus</em><span> is 2.0–2.7 ‘X’ – a difference that is not difficult to estimate. An additional new approach suggested now, particularly for L</span>₃<span> of species and/or genera difficult to differentiate (such as </span><em>Chabertia ovina</em><span> and </span><em>Oesophagostomum columbianum</em><span>), is to estimate the proportion of the larval sheath tail extension comprising a terminal thin, whip-like filament. For the experienced person, it is seldom necessary to measure more than one or two sheath tail extensions of L</span>₃<span> in a mixed culture, because the identity of most of the remaining L</span>₃<span> can thereafter be estimated in relation to those measured, without having to take further measurements. The aim of this article was to present the novel approach in the form of a working guide for routine use in the laboratory. To facilitate identification, figures and a separate organogram for each of small ruminants and cattle have been added to illustrate the distinguishing features of the common L</span>₃<span>.</span><br />

In 2004, a new concept was introduced for simplifying identification of larvae of the common nematodes of cattle, sheep and goats that comprises estimates of the lengths of the sheath tail extensions of infective third-stage larvae (L3) of each genus and/or species to that of Trichostrongylus spp., instead of having to be dependent only on measurements in micrometre. For example, if the mean length of the sheath tail extension (the extension of the sheath caudad, beyond the caudal tip of the larva) of Trichostrongylus colubriformis and Trichostrongylus axei is assumed to be ‘X’, then that ofHaemonchus contortus is 2.0–2.7 ‘X’ – a difference that is not difficult to estimate. An additional new approach suggested now, particularly for L3 of species and/or genera difficult to differentiate (such as Chabertia ovina and Oesophagostomum columbianum), is to estimate the proportion of the larval sheath tail extension comprising a terminal thin, whip-like filament. For the experienced person, it is seldom necessary to measure more than one or two sheath tail extensions of L3 in a mixed culture, because the identity of most of the remaining L3 can thereafter be estimated in relation to those measured, without having to take further measurements. The aim of this article was to present the novel approach in the form of a working guide for routine use in the laboratory. To facilitate identification, figures and a separate organogram for each of small ruminants and cattle have been added to illustrate the distinguishing features of the common L3.