Analysis of milk leukocyte differential measures for use in management practices for decreased mastitis incidence.

The aim of this study was to assess the usefulness of measures derived from milk leukocyte differential (MLD) in practices that improve fresh cow mastitis monitoring and decrease mastitis incidence. Quarter milk samples were collected from Holstein and Jersey cows on d 4 and 11 postcalving. Samples were analyzed using MLD, whereby cell counts and quarter infection diagnosis were obtained. Measures derived from MLD included cell scores (total leukocyte, neutrophil, macrophage, and lymphocyte scores), cell proportions (neutrophil, macrophage, and lymphocyte percentages), cell thresholds (total leukocyte, neutrophil, macrophage, and lymphocyte thresholds), and MLD diagnosis at different threshold settings (A, B, and C). Microbiological culturing of milk samples was used to determine infection status to compare the MLD diagnosis and serve as an indicator of infection. Measures derived from the microbiological analysis included occurrence of major pathogens, minor pathogens, and infection. Data analysis was based on a linear mixed model, which was used on all measures for the estimation of the fixed effects of breed, lactation number, day of sample collection, time of sampling, and quarter location, and the random effects of animal and week of sampling. All the fixed effects studied were significant for one or more of the analyzed measures. The results of this study showed that MLD-derived measures justify further study on their use for management practices for mastitis screening and prevention in early lactation.

Immune-mediated responses account for the majority of production loss for grazing meat-breed lambs during Trichostrongylus colubriformis infection.

The hypothesis tested in this experiment was that Trichostrongylus colubriformis infection would reduce growth rates of grazing meat-breed lambs; however production loss would be reduced by suppression of the host immune response. The experiment had a 3×2 factorial design using 6-7 month old meat-breed lambs which remained uninfected or infected (IFY) with 2000 or 4000 T. colubriformis L3/week for 12 weeks and were immunosuppressed (SUPY) using methylprednisolone acetate once weekly or remained non-immunosuppressed (SUPN). Immunosuppression increased worm egg counts (WEC) of infected lambs (SUPY 2421 eggs per gram (epg), SUPN 1154 epg on day 84, p<0.05) and T. colubriformis burdens (p<0.05-0.10) and reduced circulating eosinophils (p<0.05 on days 11, 42, 56 and 84) and intestinal total antibody titres (p<0.02). There was a significant (p<0.05) interaction between the main effects of infection and immunosuppression with infection having a larger negative effect on the liveweight of non-immunosuppressed lambs. The immunological response of the host to T. colubriformis infection accounted for 75% of the overall cost of infection (3.1kg) with the majority of this cost occurring during the first 35 days of infection. In contrast, most of the cost associated with the direct effect of infection occurred after day 35. These results confirm in grazing meat-breed lambs that the host's immunological response to T. colubriformis infection is the major component of production loss.

Stress assessment in captive greylag geese (Anser anser).

Chronic stress--or, more appropriately, "allostatic overload"--may be physiologically harmful and can cause death in the most severe cases. Animals in captivity are thought to be particularly vulnerable to allostatic overload due to artificial housing and group makeup. Here we attempted to determine if captive greylag geese (Anser anser), housed lifelong in captivity, showed elevated levels of immunoreactive corticosterone metabolites (CORT) and ectoparasites in dropping samples as well as some hematological parameters (hematocrit, packed cell volume, total white blood cell count [TWBC], and heterophil:lymphocyte ratio [H:L]). All of these have been measured as indicators of chronic stress. Furthermore, we correlated the various stress parameters within individuals. Captive geese showed elevated values of CORT and ectoparasites relative to a wild population sampled in the vicinity of the area where the captive flock is held. The elevated levels, however, were by no means at a pathological level and fall well into the range of other published values in wild greylag geese. We found no correlations between any of the variables measured from droppings with any of the ones collected from blood. Among the blood parameters, only the H:L negatively correlated with TWBC. We examine the problem of inferring allostatic overload when measuring only 1 stress parameter, as there is no consistency between various measurements taken. We discuss the different aspects of each of the parameters measured and the extensive individual variation in response to stress as well as the timing at which different systems respond to a stressor and what is actually measured at the time of data collection. We conclude that measuring only 1 stress parameter often is insufficient to evaluate the well-being of both wild and captively housed animals and that collecting behavioral data on stress might be a suitable addition.

Assessment of exercise-induced alterations in neutrophil function in horses.

OBJECTIVE: To evaluate the effects of a standardized exercise test to exhaustion in horses on leukocyte function ex vivo. ANIMALS: 6 Thoroughbred geldings. PROCEDURES: Blood samples were obtained from each horse before exercise; at exhaustion (termed failure); and at 2, 6, 24, 48, and 72 hours after exercise to evaluate hematologic changes, rate of leukocyte apoptosis, and leukocyte production of reactive oxygen species (ROS) ex vivo. To assess leukocyte function, leukocyte ROS production in response to stimulation with lipopolysaccharide, peptidoglycan, zymosan, and phorbol myristate acetate was evaluated. Apoptosis was evaluated via assessment of caspase activity in leukocyte lysates. RESULTS: In response to lipopolysaccharide, production of ROS by leukocytes was significantly increased at 2 hours and remained increased (albeit not significantly) at 6 hours after exercise, compared with the preexercise value. In the absence of any stimulus, leukocyte ROS production was significantly increased at 6 and 24 hours after exercise. In contrast, ROS production in response to phorbol myristate acetate was significantly decreased at 6, 24, and 72 hours after exercise. Leukocyte ROS production induced by zymosan or peptidoglycan was not altered by exercise. Leukocytosis was evident for 24 hours after exercise, and neutrophilia was detected during the first 6 hours. A significant increase in the rate of leukocyte apoptosis was detected at failure and 72 hours after exercise. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that strenuous exercise undertaken by horses causes alterations in innate immune system functions, some of which persist for as long as 72 hours after exercise.

A standardized cytological and immunochemical method for the analysis of fine-needle spleen aspirates: assessment of leukocyte population changes in canine visceral leishmaniosis.

A method for the evaluation of splenic cellularity using samples collected by fine-needle aspirative biopsy was standardized in this work. The procedure includes erythrocyte lysing, preparation of cytospin films and staining by histochemical and immunocytochemical techniques. The cellular profiles of spleen preparations were compared with those observed in peripheral blood samples subjected to the same procedure. Two groups were compared, one consisting of 14 healthy uninfected and the other of 15 polysymptomatic Leishmania chagasi/infantum-infected dogs, from an endemic area for visceral leishmaniosis. Cell populations were identified by conventional hematoxilin-eosin and Wright' stainings, and by immunocytochemistry using monoclonal antibodies against canine CD45RA and CD45RB, phagocytes and a pan-leukocyte antigen. Larger neutrophil (P < 0.0001) and monocyte/macrophage (P = 0.0036) relative counts and lower lymphocyte relative counts (P < 0.0001) were found in the spleen, and not in the blood, of the animals with leishmaniosis than in those of the healthy animals. The proportions of CD45RB+ cells were higher, and of CD45RA+ cells were lower, both in the spleen and in the blood of animals with leishmaniosis than in those of healthy dogs (P < 0.05). Additionally, hematoxilin-eosin-stained cytospins of spleen aspirates from Leishmania-infected animals permitted the easy visualization of amastigote forms inside phagocytes, under light microscopy.

Improving resilience against natural gastrointestinal nematode infections in browsing kids during the dry season in tropical Mexico.

The objective was to determine the effect of supplementary feeding on the resilience and resistance of Criollo kids against natural gastrointestinal nematode (GIN) infections, when browsing native vegetation during the dry season in tropical Mexico. Thirty-three two-month-old Criollo kids, raised nematode free, were included at weaning in a 20-week trial. The kids were placed into four groups. Two groups of eight kids were offered 100g/day soybean and sorghum meal (26%:74% respectively fresh basis) (treated/supplemented (T-S) and infected/supplemented (I-S)). Two groups remained with no supplement for the duration of the trial (infected/non-supplemented (I-NS) (n=9) and treated/non-supplemented (T-NS) (n=8)). Kids in groups T-S and T-NS were drenched with 0.2mg of moxidectin/kg body weight orally (Cydectin, Fort Dodge) every 28 days. Groups I-S and I-NS were naturally infected with GIN. The animals browsed native vegetation for an average of 7h/day together with a herd of 120 naturally infected adult goats. Cumulative live weight gain (CLWG), packed cell volume (PCV), haemoglobin (Hb), total plasma protein and plasma albumin were recorded every 14 days as measurements of resilience. Resistance parameters (faecal egg counts (FEC) and peripheral eosinophil counts (PEC)) were also measured. Bulk faecal cultures were made for each group every 28 days. Every month a new pair of initially worm-free tracer kids assessed the infectivity of the vegetation browsed by the animals. Tracer kids and faecal cultures showed that kids faced low mixed infections (Haemonchus contortus, Trichostrongylus colubriformis and Oesophagostomum columbianum). Under conditions of scarce vegetation, such as those in the present study, supplemented groups (I-S and T-S) had higher growth rates compared to the non-supplemented groups independently of the control of GIN infection with anthelmintic (AH) treatment (P<0.001). Supplementary feeding did not affect FEC or PEC. In the absence of supplementation, lack of AH treatment may lead to outbreaks of clinical nematodosis. The supplementary feeding was economically feasible.

The effectiveness of classical swine fever surveillance programmes in The Netherlands.

Consequences of classical swine fever (CSF) epidemics depend on the control measures, but also on the number of infected herds at the end of the high-risk period (HRP). Surveillance programmes aim to keep this number as low as possible, so the effectiveness of surveillance programmes can be measured by the number of infected herds at the end of the HRP. In this paper, an evaluation of the effectiveness of the following five Dutch CSF surveillance programmes is presented: (1) routine gross pathology of severely diseased pigs; (2) routine virological tests of tonsils of all pigs, submitted under 1; (3) daily clinical observation by the farmer; (4) periodic clinical inspection by a veterinarian; (5) leucocyte counts in blood samples from diseased animals on a herd where antimicrobial 'group therapy' is started. The evaluation was done by a modelling study, in which virus transmission, disease development, and actions and diagnostic tests in surveillance programmes were simulated. Also, the yearly costs of the programmes were calculated, and direct costs of CSF epidemics were related to the number of infected herds at the end of the HRP. It appeared that the current Dutch surveillance programmes, without the leucocyte counts, keep the number of infected herds at the end of the HRP below 20 with 95% probability. Leaving out the most-expensive programme of periodic inspection (12.5M per year) does not change this result - indicating that (for CSF surveillance) the programme could well be stopped. If the leucocyte programme, which is currently not effective due to the low sample submission rate, optimally were applied, the 95th percentile could be reduced to 10 infected herds. However, whether application is beneficial is unclear, because of uncertainty of the economic benefits due to the many expected false-positive herds each year.

Assessment of respiratory herd health in weaner pigs by measuring cellular composition of bronchoalveolar lavage fluid.

Cell differentiation of bronchoalveolar lavage fluid (BALF) was compared among 11 herds having a history of recurrent respiratory disease in weaner pigs and nine herds lacking such a history. In every herd, 20 pigs aged 8-10 weeks were lavaged. The two groups differed significantly on median percentage of macrophages, neutrophils and lymphocytes, but not on white blood cell count of the BALF. Logistic regression showed the percentage of samples per herd exceeding the reference value for neutrophils of 0-8% to be the most promising parameter to assess the health status in weaner pigs.

Comparative assessment of the erythrocyte osmotic fragility and of haematological and plasma biochemical values in the Nigerian white Fulani and N'dama breeds of cattle.

The blood profiles of the Nigerian White Fulani and N'dama breeds of cattle were compared. The White Fulani cattle had a significantly higher haemoglobin concentration (p < 0.001), mean corpuscular haemoglobin (p < 0.05), plasma sodium (p < 0.02), total protein (p < 0.01), albumin (p < 0.01) and globulin (p < 0.02), but lower neutrophil counts (p < 0.01) and osmotic fragility of erythrocytes than the N'dama cattle. The plasma potassium, calcium, bicarbonate, inorganic phosphate, albumin/globulin ratio, urea, creatinine, packed cell volume and mean corpuscular volume, and the erythrocyte, leukocyte, lymphocyte, eosinophil and monocyte counts were similar in the two breeds.

Clinical outcome and diseases associated with extreme neutrophilic leukocytosis in cats: 104 cases (1991-1999).

OBJECTIVE: To describe diseases, prognosis, and clinical outcomes associated with extreme neutrophilic leukocytosis in cats. DESIGN: Retrospective study. ANIMALS: 104 cats with extreme neutrophilic leukocytosis. PROCEDURE: Medical records from 1991 to 1999 were examined to identify cats that had > or =50,000 WBC/microl with > or =50% neutrophils. Signalment, absolute and differential WBC counts, rectal temperature, clinical or pathologic diagnosis, duration and cost of hospitalization, and survival time were reviewed. RESULTS: Mean age of cats was 8.3 years, mean WBC count was 73,055 cells/microl, and mean absolute neutrophil count was 59,046 cells/microl. Mean duration of hospitalization was 5.9 days, and mean cost of hospitalization was $2,010. Twenty-nine (28%) cats were febrile, and 63 (61%) cats died. Overall median survival time was 30 days. Cats with neoplasia were nearly 14 times as likely to die unexpectedly as cats with other diseases. CONCLUSIONS AND CLINICAL RELEVANCE: Extreme neutrophilic leukocytosis was associated with a high mortality rate. The prognostic importance of extreme neutrophilic leukocytosis should not be overlooked. Cats and dogs have similar diseases, mortality rates, and treatment costs associated with extreme neutrophilic leukocytosis.

[The leukocyte count is a valuable parameter for detecting classical swine fever]. / Leukocytentelling is een waardevol hulpmiddel voor het opsporen van klassieke varkenspest.

In this paper we describe a study of the use of the white blood cell count (wbcc) as a parameter for detecting outbreaks of Classical Swine Fever (CSF). Meta-analysis of the results of challenge experiments revealed that oronasal infection of SPF-pigs with the virulent CSF virus (CSFV) strains Brescia or NL9201 resulted in a significant decrease in the average white blood cell count during the first week after inoculation of the virus. Challenge of conventional finishing pigs and sows with the moderately virulent strain Paderborn also resulted in a significant decrease in the average wbcc. However, this decrease was not observed after inoculation of SPF pigs with the mildly virulent CSFV strains Henken, Zoelen, or Bergen. The usefulness of clinical inspection in combination with wbcc to detect CSF outbreaks in the field was examined using the results of 214 EDTA blood specimens collected from 22 infected herds and 7250 EDTA blood specimens collected from 1450 non-infected herds. Half of the infected herds had been infected with the moderately virulent CSFV strain Venhorst (closely related to strain Paderborn) during the 1997-98 epidemic in the Netherlands. The other half had been infected with the moderately virulent CSFV strain Loraine. Using these data as a starting point, 1000 samples of one to ten specimens were generated by Monte Carlo simulation. These simulated samples and the samples of the non-infected herds were analysed by use of Receiver Operating Characteristic curves. On the basis of that analysis, the optimal number of animals whose wbcc needed to be determined to detect a CSF outbreak was five. With this number of animals, in conjunction with the threshold of 8000 white blood cells per mm3 (meaning that a herd is designated as CSF suspect if one or more of the five specimens has a white blood cell count of 8000 leukocytes/mm3 or less), the test procedure had a herd sensitivity (HSE) of 94.5% and a herd specificity (HSP) of 97.2%). The HSE is defined as the percentage of samples of infected herds with a positive result of the test procedure; HSP is defined as the percentage of uninfected herds with a negative result of the test procedure. We conclude that the wbcc can help the veterinary practitioner to detect outbreaks of CSF caused by (moderately) virulent CSFV strains. However, for the detection of outbreaks caused by mildly virulent CSFV strains, the contribution of the wbcc is doubtful. Development of additional tools that can improve the clinical diagnosis of the veterinary practitioner remains desirable.

Clinical outcome and associated diseases in dogs with leukocytosis and neutrophilia: 118 cases (1996-1998)

OBJECTIVE: To describe diseases, prognosis, and clinical outcomes associated with leukocytosis and neutrophilia in dogs. DESIGN: Retrospective study. ANIMALS: 118 dogs with leukocytosis and neutrophilia. PROCEDURE: Medical records from 1996 to 1998 were examined for dogs with WBC > or = 50,000 cells/microliter and neutrophilia > or = 50%. Signalment, absolute and differential WBC counts, body temperature, clinical or pathologic diagnosis, duration and cost of hospitalization, and survival time were reviewed. RESULTS: Mean age was 7.7 years, WBC count was 65,795 cells/microliter, and absolute neutrophil count was 53,798 cells/microliter. Mean duration of hospitalization was 7.4 days and cost of hospitalization was $2,028.00. Forty (34%) dogs were febrile, and 73 (62%) dogs died. Overall median survival time was 17 days. Dogs with neoplasia or fever were more likely to die than dogs that were hospitalized or had systemic or local infections. CLINICAL IMPLICATIONS: Leukocytosis and neutrophilia were associated with high mortality rate and have prognostic value. Given the mean duration and cost of hospitalization, frank discussion with an owner at first recognition of leukocytosis and neutrophilia may be warranted.

Assessment of phagocytic capacity and opsonic activity in blood and mammary secretions during lactation in sows.

The objective of this study was to determine whether phagocytic capacity and opsonic activity in blood and mammary secretions of sows are impaired at parturition compared with later on during lactation. The study comprised eight primiparous sows (Landrace x Yorkshire) free from clinical signs of disease. Blood and mammary secretion samples were collected within 48 h of parturition and 7 and 16 days after parturition. Numbers and proportion of polymorphonuclear neutrophils (PMN) were determined in blood and mammary secretions. Phagocytic capacity was assessed in whole blood and in a cell suspension derived from mammary secretions. Opsonic activity was assessed in serum and i cell-depleted, skimmed mammary secretions. The two assays were based on chemiluminescence, both having zymosan and Escherichia coli as target particles. Numbers and proportion of PMN in mammary secretions were higher (P < 0.05) at parturition than later on during lactation. A parturition, phagocytic capacity in cell suspensions derived from mammary secretions was higher for both (P < 0.05) and E. coli (P < 0.1). However, when phagocytic capacity was related to the number of PMN in the suspension no such difference was observed. The opsonic activity in cell-depleted, skimmed mammary secretions at parturition was lower (P < 0.05) for zymosan but not for E. coli. None of the described variations were reflected in blood or serum. The findings of this study do not unequivocally support the theory that an immune suppression at parturition in the sow can help explain the increased incidence of coliform mastitis at that time.

[The assessment of the status of bottlenose dolphins Tursiops truncatus in the period after their capture]. / Otsenka sostoianiia del'finov afalin Tursiops truncatus v period posle pleneniia.

Blood (eosinophil count and glucose content) and heart rate parameters were studied for assessment of individual responses and level of stress reaction of recently captured bottle-nosed dolphins Tursiops truncatus. Cross-tabulation analysis was carried out of the physiological parameters and the following process of adaptation to captivity. The normal level of stress response in contrast to excessive or abnormal responses were of the best predictive value for adaptation and survival of the animals. Correlation was found between the blood system parameters and heart rate variability in stress response assessment.

Clinical and clinicopathological assessment of serial phlebotomy in the Sprague Dawley rat.

Two studies, designed to mimic a single-dose, cross-over pharmacokinetic protocol, were conducted to gain a better understanding of the rat's response to multiple, frequent blood sampling. Parameters evaluated included body weight, clinical signs of disease, hematologic and serum biochemical analytes, organ weights, and histopathologic features. Study groups consisted of either 6 or 8 male, viral antibody-free, Sprague Dawley rats. These included controls and blood-collection groups that represented withdrawal of 10, 15, 20, 30, and 40% of estimated total blood volume. Volume of blood collected per time point was based on the total volume to be withdrawn divided by the 13 samples that were collected over 24 h. This regimen was repeated 2 weeks later. Samples were taken for clinical pathologic evaluation on the days subsequent to blood collection for both studies as follows: 0, 1, 2, and 3 days; 7, 8, or 9 days; and either 13 or 14 days. In Study 1, samples were also taken on either days 15 or 16, and on 17 or 18 after the second blood collection. Approximately 2 weeks after the second blood collection regimen, animals were euthanized. Animals in one study were necropsied, and selected tissues were taken for histologic examination. Analysis of variance, based on changes from baseline, was used to assess group differences. Results indicate that the rate of body-weight gain for the bled groups was not significantly different from that of the controls. Group differences in multiple hematologic parameters were significant. Changes were typical of acute blood-loss anemia, with positive or negative trends relating to the volume of blood removed. In addition, these changes were characterized by recovery to control values within approximately 14 days. Few statistically significant group differences were detected in serum biochemical values, and those detected were not biologically relevant. Although organ weights of bled groups were similar to those of controls, minimal to mild splenic hematopoiesis was present in all bled groups, compared with controls. These data indicate that removal of up to 40% of a rat's total blood volume over a 24-h period, and repeated 2 weeks later, caused no gross ill effects.

In vitro assessment of the influence of different leucocyte count lavaged from normal and inflamed cow uterus on the mouse embryo development.

The aim of this study was to evaluate the effect of different leucocyte count from normal and inflamed uterus in excess of PMNs on the development of mouse embryos during the morula or blastocyst stages in vitro. Results showed that cultivation of blastocysts in the presence of leucocytes washed from the uterus, after mediation of specific or nonspecific endometritis at a concentration of 1 x 10(4), led to significant inhibition of embryo development. No similar observations were made with blastocysts cultivated in the presence of leucocytes obtained from the normal uterus at 2-4 days after spontaneous ovulation.

Feline infectious peritonitis: a review of clinicopathological changes in 65 cases, and a critical assessment of their diagnostic value.

In 65 natural cases of feline infectious peritonitis (FIP) the common clinicopathological changes included lymphopenia (77 per cent), neutrophilia (45 per cent), anaemia (37 per cent), hyperproteinaemia (39 per cent) and hyperglobulinaemia (39 per cent). There was no difference in the frequency of these abnormalities between the 38 cases of effusive disease and the 27 cases of non-effusive disease. The most consistent changes shown by serum protein electrophoresis were increases in alpha 2- and gamma-globulins. The protein content of the effusions ranged from 39 to 98 g/litre with the globulins comprising 50 to 82 per cent. Coronavirus serology showed a wide variation in antibody titres (0 to 2560) with 320 the modal titre. The diagnostic value of this information was evaluated by comparing it with data from 65 cats in which FIP was considered as a differential diagnosis, but another disease was diagnosed. None of the laboratory tests, including coronavirus serology, had good sensitivity and specificity for the diagnosis of the disease. The presence of multiple abnormalities compatible with the disease increased the specificity but decreased the sensitivity of the diagnosis.