Michael P. Martin

Reliability and validity of a modified gait scoring system and its use in assessing tibial dyschondroplasia in broilers

1. The gait scoring system for broilers developed by Kestin et al . ( Veterinary Record , 131: 190-194, 1992) has been widely used to evaluate leg problems. The many factors and measures associated with this scale have empirically established its external (biological) validity. However, published test-retest (within-observer) reliabilities are poor, and inter-observer reliabilities are unknown. We evaluated several modifications to this scale aimed at improving its objectivity and reliability. 2. Eighteen naïve observers scored a standardised video of birds exhibiting varying degrees of lameness, either using Kestin et al .s system, or our modified system. 3. Test-retest reliability (0.906) for Kestin et al .s system was higher than previously reported. Interrater reliability was also good (0.892). The modified system offered significantly better test-retest (0.948) and inter-rater reliabilities (0.943), without incurring costs in terms of time taken or difficulty of use. The systems were consistent, assigning individual birds the same score on average. 4. It is concluded that the modified system offers the advantages of reduced error within and between studies. 5. In a second experiment, we used our modified scoring system to examine the relationship between tibial dyschondroplasia (TD) and gait score in 267 selected broilers. 6. Neither the presence nor severity of TD affected gait score, suggesting that, at least in this strain of broilers, other leg problems like slipped tendons or torsional deformities had more influence on gait impairment than did TD.

Experimental Reproduction of Enterococcal Spondylitis in Male Broiler Breeder Chickens

Abstract There has been a recent emergence of epidemic spinal infections with necrosis causing lameness and mortality in male broilers and broiler breeders. Mortality in affected flocks may be as high as 15%. The disease has been called enterococcal spondylitis (ES), based on the frequent isolation of Enterococcus cecorum from the lesions and necrosis and inflammation observed in the free thoracic vertebrae (FTV) of affected birds. Male broiler breeders in an experimental setting were challenged with pure E. cecorum isolates obtained from ES-affected commercial flocks. Challenge routes included oral gavage (108), intravenous (IV; 103), and air sac (AS; 103). Half the study birds in each group were chemically immunosuppressed with dexamethasone. Spinal lesions were observed grossly in birds challenged intravenously (2.9%) and birds challenged orally (6.1%). Microscopic spinal lesions consistent with ES were more frequently identified compared with gross lesions in the orally challenged group (30.3%). Chemical immunosuppression with dexamethasone was not associated with a greater incidence of ES in this study. By recreating the disease experimentally, the study design reported here may help in the further development of an experimental challenge model for future studies on risk factors, prevention, and therapeutic intervention of ES.

An Outbreak and Source Investigation of Enterococcal Spondylitis in Broilers Caused by Enterococcus cecorum

SUMMARY. Enterococcus cecorum was isolated from spondylitis lesions in broilers from two flocks in North Carolina that were experiencing increased mortality. Affected birds showed paresis and paralysis, clinical signs characteristic of enterococcal spondylitis (ES). Affected birds rested on their hocks and caudal abdomens with legs extended forward and were unable to stand or walk. Necropsy examination of affected birds revealed firm to hard inflammatory masses involving the vertebral bodies at the level of the free thoracic vertebra that bulged dorsally and compressed the spinal cord. When opened, lesions contained pale, tan to yellow caseonecrotic material. Microscopically, necrosis and fibrinoheterophilic spondylitis with intralesional gram-positive bacteria were seen. Heavy growth of E. cecorum recovered from vertebral lesions confirmed the diagnosis of ES. To investigate possible sources of the organism for one of the flocks bacterial cultures were made from the environment, water lines, mice trapped on the farm, cecal/cloacal swabs from one of the parent broiler breeder flocks, egg residue, hatching eggs, and the hatchery environment. Except for cecal/cloacal swabs from the breeders, E. cecorum was not isolated from any of these samples. When compared phenotypically and genotypically, cecal/cloacal isolates of E. cecorum from the breeders differed from isolates from spondylitis lesions in the broilers. The source of E. cecorum for the broiler flocks was not determined, but vertical transmission appears unlikely.

Selected Blood Chemistry and Gas Reference Ranges for Broiler Breeders Using the i-STAT® Handheld Clinical Analyzer

Abstract Selected blood chemistry and gas reference ranges for clinically healthy broiler breeder hens were established using CG8+ cartridges in an i-STAT® handheld point-of-care clinical analyzer. Samples from 165 hens (25–36 wk of age), representing three broiler breeder strains reared by four integrators, were evaluated. A standardized sampling technique was developed to minimize instrument error readings. The following reference ranges and means, respectively, were determined: sodium (141.6–152.6, 147.1 [mmol/L]), potassium (4.1–5.7, 4.9 [mmol/L]), ionized calcium (1.20–1.73, 1.47 [mmol/L]), glucose (207.2–260.7, 234.0 [mg/dl]), hematocrit (21.3–30.8, 26.1 [% packed cell volume]), hemoglobin (7.3–10.5, 8.9 [g/dl]), pH (7.28–7.57, 7.42), carbon dioxide partial pressure (25.9–49.5, 37.7 [mm Hg]), oxygen partial pressure (32.0–60.5, 46.2 [mm Hg]), bicarbonate (18.9–30.3, 24.6 [mmol/L]), total carbon dioxide (19.9–31.5, 25.7 [mmol/L]), base excess (−6.8 to 7.2, 0.2), and oxygen saturation (70.6–93.3, 82.0 [%]). Wide ranges in blood gases and base excess occurred in all strains. Cobb strain hens had significantly lower glucose and higher partial and saturated oxygen values compared with two Ross strains. Significant differences in several blood parameters were found among different integrators and in older postpeak production birds. The i-STAT handheld point-of-care clinical analyzer provides rapid, relatively low cost, blood chemistry values that are useful for investigating broiler breeder flock diseases of unknown or uncertain etiology, especially those suspected of having a metabolic cause.

Molecular epidemiology of Enterococcus cecorum isolates recovered from enterococcal spondylitis outbreaks in the southeastern United States

Enterococcus cecorum, a normal intestinal inhabitant, is increasingly responsible for outbreaks of arthritis and osteomyelitis in chickens worldwide. Enterococcal spondylitis (ES) is a specific manifestation of E. cecorum-associated disease in which increased flock morbidity and mortality result from chronic infection involving the free thoracic vertebra. In this study the genetic relatedness and antimicrobial resistance of isolates recovered from ES-affected flocks in the southeastern United States were determined. ES outbreaks from 2007 to 2011 were investigated in North Carolina (15 flocks, 13 farms, four integrators), South Carolina (one flock, one farm, one integrator) and Alabama (six flocks, six farms, one integrator). From these 22 epidemiologically distinct outbreaks, 326 isolates of E. cecorum were recovered. Isolates from spinal lesions and caeca of affected birds (cases) and caeca of unaffected birds (controls) were genotyped using pulsed-field gel electrophoresis; phenotyped using both GenIII MicroPlate™ (Biolog; Hayward, CA, USA) microbial identification plates and antimicrobial sensitivity testing; and compared with each other. Isolates from spinal lesions were incapable of mannitol metabolism and the majority of these isolates were genetically clonal. In contrast, caecal isolates from control birds varied in their ability to metabolize mannitol and were genetically diverse. Isolates from both case and control birds had high levels of antimicrobial resistance. These findings indicate that the increase in E. cecorum-associated disease in the southeast United States is due to the emergence of new clones with increased pathogenicity and multidrug resistance.

Disseminated Aspergillus flavus Infection in Broiler Breeder Pullets

Abstract Increased morbidity and mortality occurred in a 5-wk-old broiler breeder replacement pullet flock. The affected broiler pullet flock was housed on the first floor of a two-story confinement building. Mortality increased to 0.1%/day compared to the flock on the second floor, which had mortality levels of less than 0.01%/day. Clinical signs in the affected chickens included inactivity, decreased response to stimuli, and anorexia. No respiratory or neurologic signs were observed. On necropsy, affected pullets were dehydrated and emaciated and had disseminated variably sized single or multiple heterophilic granulomas that contained intralesional septate and branching fungal hyphae. Lesions were extensive around the base of the heart in the thoracic inlet and in the kidneys. Other affected organs included eyelid, muscle, proventriculus, ventriculus, intestine, liver, spleen, lung, and heart. Aspergillus flavus was cultured from the visceral granulomas. The source of flock exposure to the organism was not determined.

Bacterial orchitis and epididymo-orchitis in broiler breeders.

Two cases of orchitis and epididymo-orchitis in broiler breeders are described. The first case occurred in a 62-week-old rooster showing signs of breeding inactivity. Grossly, the left testis was abnormally shaped and had multiple yellow foci, prominent blood vessels, and a gelatinous mass on the surface. The right testis and other tissues appeared normal. A pure, heavy growth of Staphylococcus aureus was obtained on bacterial culture, and intralesional Gram-positive cocci were numerous on histopathology. The second case occurred in a 28-week-old rooster that was found dead. Both testes were abnormally shaped, swollen, firm, and had irregular, dark, depressed areas, prominent vessels, and multiple petechial haemorrhages. Epididymides were enlarged and nodular. Other lesions in this rooster included poor nutritional condition, distended cloaca, and urate scalding of the abdominal skin below the vent. Microscopically, there was extensive, severe heterophilic intratubular orchitis and epididymitis with intralesional Gram-negative bacteria. A heavy, pure growth of Escherichia coli was obtained from the testes and epididymides. In both cases, the findings suggested that the most probable route of infection was ascending via the ductus deferens.

Pathogenesis of Enterococcal Spondylitis Caused by Enterococcus cecorum in Broiler Chickens

Enterococcal spondylitis (ES) is a disease of commercial broiler chickens, with a worldwide distribution. Symmetrical hind limb paralysis typical of ES results from infection of the free thoracic vertebra (FTV) by pathogenic strains of Enterococcus cecorum . To determine the pathogenesis of ES, birds with natural and experimental ES were studied over time. In natural disease, case birds (n = 150) from an affected farm and control birds (n = 100) from an unaffected farm were evaluated at weeks 1–6. In control birds, intestinal colonization by E. cecorum began at week 3. In case birds, E. cecorum was detected in intestine and spleen at week 1, followed by infection of the FTV beginning at week 3. E. cecorum isolates recovered from intestine, spleen, and FTV of case birds had matching genotypes, confirming that intestinal colonization with pathogenic strains precedes bacteremia and infection of the FTV. Clinical intestinal disease was not required for E. cecorum bacteremia. In 1- to 3-week-old case birds, pathogenic E. cecorum was observed within osteochondrosis dissecans (OCD) lesions in the FTV. To determine whether OCD of the FTV was a risk factor for ES, 214 birds were orally infected with E. cecorum, and the FTV was evaluated histologically at weeks 1–7. Birds without cartilage clefts of OCD in the FTV did not develop ES; while birds with OCD scores ≥3 were susceptible to lesion development. These findings suggest that intestinal colonization, bacteremia, and OCD of the FTV in early life are crucial to the pathogenesis of ES.

Evaluation of the Effectiveness of Two Infectious Bronchitis Virus Vaccine Programs for Preventing Disease Caused by a California IBV Field Isolate

Abstract Infectious bronchitis virus CA99 serotype was isolated from several broiler flocks in Northern California. The virus caused late-onset respiratory disease and increased airsacculitis condemnation in affected flocks despite the use of an established infectious bronchitis virus vaccination program. An experimental study compared Holland/Arkansas and Massachusetts/Arkansas vaccination protocols to determine the efficacy of commercial infectious bronchitis virus vaccines in reducing respiratory disease and airsacculitis lesions found at processing that were associated with a CA99 field isolate. All vaccination groups were given Massachusetts/Connecticut strains of infectious bronchitis virus vaccines at age 1 day followed by vaccination with either Holland/Arkansas or Massachusetts/Arkansas vaccine strains at 18 days of age. Birds were challenged at age 31 days with a CA99 field isolate. Gross pathology, histopathology, and virus isolation were evaluated. Chickens vaccinated with Holland/Arkansas had marginally better protection against CA99 challenge than chickens vaccinated with Massachusetts/Arkansas, although differences were not statistically significant.

Selected blood chemistry values in mobility-impaired broiler breeder hens with suspected calcium tetany using the i-STAT handheld clinical analyzer.

SUMMARY. Calcium tetany is a poorly defined disease of broiler breeder hens that results from acute hypocalcemia. It is characterized by impaired mobility, increased mortality, and absence of gross lesions that would explain the impaired mobility. To evaluate if hens with impaired mobility had calcium tetany or other abnormalities, blood values from normal and affected hens were determined using the i-STAT® handheld clinical analyzer. Three flocks were evaluated weekly prior to peak production (range 25–30 wk of age) comparing normal hens to hens with clinically apparent calcium tetany. Calcium tetany suspect (CaTS) hens from four additional flocks were also evaluated. Significant hypocalcemia (P < 0.001) was observed in CaTS hens (average  =  1.14 mmol/L ionized calcium [iCa]) compared to normal hens (average  =  1.53 mmol/L iCa) in only one of three flocks sampled weekly. Clinically affected hens from one of the other four flocks also had hypocalcemia. Blood value abnormalities in mobility-impaired hens without hypocalcemia included hypernatremia. Findings in this study indicate calcium tetany is one cause of impaired mobility in breeder hens, but mobility impairment without hypocalcemia can also occur. Calcium tetany should be confirmed by finding significantly decreased levels of iCa in the blood, as diagnosis based on clinical presentation and necropsy results can be inaccurate. The i-STAT® handheld clinical analyzer is an efficient, relatively low-cost method to determine iCa and other blood chemistry values that may be associated with impaired mobility in broiler breeder hens.