Enzootic calcinosis in Toggenburg goats in New Zealand.

CASE HISTORY: Necropsies on Toggenburg goats culled from a small farm in the Manawatu district of New Zealand, performed at Massey University (Palmerston North, NZ) over a period of 29 years (1991-2019), revealed soft tissue mineralisation, particularly of cardiovascular tissues. The farm spans 10 acres and runs between 15 and 30 Toggenburg goats. The goats are predominantly on pasture comprising a variety of types. PATHOLOGICAL FINDINGS: Necropsies were performed on all adult goats (n = 45) that died or were euthanised. Histopathology was performed on 42 goats (93%), of which 33 (73%) included sufficient tissues diagnostically relevant to soft tissue mineralisation. The most significant gross findings were in various arteries, with the aorta most commonly affected, followed by the heart and lungs. The aortic intima showed prominent, multifocal to coalescing, raised, wrinkled, white plaques. Microscopically there were multiphasic lesions of mineralisation, chondroid, and osseous metaplasia in the elastic arteries, aorta, heart and lungs. A lumbar vertebra from one goat had prominent, basophilic, fibrillar, tangled matrix lining Haversian canals and lamellae. LABORATORY FINDINGS: Blood samples were collected from 15 adult goats in the affected herd and from 10 adult Toggenburg goats from an unaffected herd. Samples were collected by jugular venipuncture at 2-month intervals for 12 months (April 2018-March 2019). Concentrations of calcium, phosphorus, 25-hydroxyvitamin D2 and D3 (25OHD2, 25OHD3) in serum were analysed. The concentration of total 25OHD in serum was 34.2 (95% CI = 18.9-49.4) nmol/L (p < 0.001) higher in goats from the affected herd than in goats from the unaffected herd. Serum 25OHD2 concentration was 46.2 (95% CI = 39.2-53.2) nmol/L higher (p < 0.001) in goats from the affected herd compared to the unaffected herd. Serum Ca concentrations in affected goats were 0.101 (95% CI = 0.005-0.196) mmol/L higher (p = 0.039) than unaffected goats, but remained within the reference range. There was no evidence of a difference in serum 25OHD3 and P concentration between the herds. VEGETATION SURVEY: All paddocks on the property were surveyed every 2 months along evenly spaced line transects, and then further traversed perpendicularly to form a grid. No known calcinogenic species were identified. Known plant sources of vitamin D identified on the farm included mushrooms (species not defined), Dactylis glomerata, lichen, pine pollen, and algae. DIAGNOSIS: Soft tissue mineralisation and enzootic calcinosis. CLINICAL RELEVANCE: Veterinarians are alerted to the possibility of either enzootic calcinosis in goats and the potential occurrence of calcinogenic plants in New Zealand; or chronic vitamin D toxicosis of non-plant origin.

Postural and gait abnormalities in a group of Romney lambs due to pain induced by severe chronic cutaneous dermatophilosis.

CASE HISTORY: Fifteen lambs from a flock of 1,000, 5-month-old Romney lambs in the Manawatu region of New Zealand, developed unusual postural and gait abnormalities during the summer of 2020. In mid-January, the lambs were grazing a raphanobrassica crop (xRaphanobrassica; Brassica oleracea x Raphanus sativus) but were removed from this crop soon after as they developed dermal lesions on the pinnae consistent with photosensitisation (colloquially known as "rape scald"). The lambs were placed onto a shaded paddock containing a pasture of predominantly perennial ryegrass (Lolium perenne) and white clover (Trifolium repens). One week following removal from the crop (late January), the 15 lambs developed abnormal postures and gaits that were described by farm staff as "neurological" and "swayback-like". CLINICAL FINDINGS: From a distance, all 15 lambs exhibited remarkably similar abnormal posture and gait whereby their weight was borne on their carpi, and they appeared to crawl while walking due to crouched hindquarters and prominent lordosis. Of the affected lambs, five were submitted for clinical examination. All five had normal vital signs. The results of neurological examinations were within normal limits. In response to palpation of the dorsum, the lambs performed an exaggerated and unusual dipping motion, particularly when pressure was applied over the lumbar spine. Parting of the fleece over the dorsum revealed thickened, indurated skin that cracked and tore easily. PATHOLOGICAL FINDINGS: The shorn pelt of a euthanised lamb revealed a large (100 cm × 40 cm), locally extensive, irregular ovoid region of the skin of the dorsum that was markedly thickened, indurated and crusted. Impression smears of the lesions revealed long filamentous chains of plump coccoid bacteria separated by transverse and longitudinal septa, consistent with Dermatophilus congolensis. Histopathology of the lesions revealed severe, chronic-active, locally extensive, necrosuppurative dermatitis with marked acanthosis, orthokeratotic and parakeratotic hyperkeratosis, ulceration, filamentous bacteria and coccoid zoospores. DIAGNOSIS: Cutaneous dermatophilosis due to Dermatophilus congolensis. CLINICAL RELEVANCE: Painful dermal lesions due to severe dermatophilosis of the dorsal skin of sheep are capable of causing postural and gait abnormalities that may be misinterpreted as signs of neurological abnormality. Such dermal lesions may be concealed by an apparently normal fleece.

Acute fibrinous pleuropneumonia and septicaemia caused by Bibersteinia trehalosi in neonatal calves in New Zealand.

Case history: In July and August 2019, 15/40, ≤48-hour-old calves became acutely ill. The calves were all born on-farm, transferred to pens soon after birth, and fed with "gold" colostrum. The hygiene, biosecurity and ventilation in the pens were poor. Of the 15 calves, 11 died or were euthanised and four calves, ≤48-hour-old, that became acutely ill later in the outbreak were treated with cefquinome, a fourth-generation cephalosporin, and recovered. Clinical findings: The affected calves presented with acute recumbency, lethargy, tachypnoea, tachycardia, increased lung sounds, inability to stand or feed, and dehydration without pyrexia. Pathological findings: Gross findings in a calf that died naturally included fibrinous pleuropneumonia, marked oedematous expansion of the interlobular septa, especially in the ventral lung lobes, fibrinous polyserositis and fibrinous polyarthritis. A second calf that was euthanised had strikingly similar lung lesions. Histologically, the pulmonary interlobular septa of both calves were prominently expanded by oedema, dilated lymphatics and the infiltration of numerous neutrophils and macrophages interspersed with small Gram-negative rod bacteria. Likewise, the visceral pleura showed fibrinopurulent inflammation with numerous small Gram-negative rods. Microbiological findings: Microbial culture and matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry identified Bibersteinia trehalosi in the lung, stifle joint and peritoneal cavity of the first calf and lung of the second. Diagnosis: B. trehalosi acute fibrinous pleuropneumonia and septicaemia. Clinical relevance: This is the first report of the clinical findings and histological lesions of B. trehalosi pleuropneumonia and septicaemia in calves in New Zealand. The pathogen is isolated with increasing frequency from cases of bovine respiratory disease in dairy cows, feedlot cattle and calves in the United Kingdom and North America. The importance of microbial culture in cases such as this with unusual lung lesions in calves <48 hours of age, cannot be over emphasised. Cefquinome was administered to all remaining heifer calves and four calves that became ill later in the outbreak recovered after cefquinome treatment.

Effect of a one-off sporidesmin challenge on the milk production of dairy cows.

AIMS To investigate the effects on milk yield in lactating dairy cows of a single dose of sporidesmin, and to categorise the responses based on clinical signs and differing degrees of liver damage, as assessed by activities of γ-glutamyl transferase (GGT) and post-mortem liver histopathology. METHODS Adult lactating dairy cows (n=17) were given a single intra-ruminal dose of 0.24 mg/kg of sporidesmin dissolved in ethanol and diluted in water on Day 0; an additional three cows served as untreated controls. Weekly serum samples were collected between Days -14 and 42 and analysed for activities of GGT. Milk yields were measured daily over the same period. Cows were subjected to euthanasia due to severe clinical signs (n=2) or were slaughtered at the end of the trial. Samples of livers were examined histologically and were scored for lesions on a scale from 0 (normal) to 3 (severe). Based on GGT activities and clinical observations, cows that were treated with sporidesmin were categorised as non-responders (no clinical signs and normal GGT), subclinical (elevated GGT and no clinical signs) or clinical. Outcomes were compared between these three groups and control cows using generalised additive models. RESULTS Seven cows were classified as clinical, and had median liver scores of 22 (95% CI=20.6-23.4), six were subclinical with median liver scores of 8.7 (95% CI=3.8-13.5) and four were non-responders with median liver scores of 2.5 (95% CI=1.2-4.3). Median liver scores for the three control cows were 1 (95% CI=-0.8-2.1). Activities of GGT increased in subclinical and clinical cows around Day 7. The milk yield of all cows treated with sporidesmin, including non-responder cows, started to decrease on Day 1, and reached a nadir (a drop of between 9 and 85%) on Day 7. CONCLUSIONS AND CLINICAL RELEVANCE It is likely that the overall effects of sporidesmin consumption on milk production by the national herd in New Zealand are hugely underestimated, especially considering its effects on non-responder and subclinical cows as shown in this trial. In view of the results presented here, the authors are suggesting a change to the definition of response to sporidesmin from non-responder, subclinical, and clinical, to subclinical-low, subclinical-high, and clinical, when measuring a combination of GGT activities, clinical signs and milk yields during facial eczema-risk seasons (summer-autumn).

Recovery of brodifacoum in vomitus following induction of emesis in dogs that had ingested rodenticide bait.

AIM: To assess the benefit of inducing emesis in dogs that have ingested rodenticide bait containing brodifacoum (BDF), by determining the amount of BDF in bait recovered from the vomitus relative to the estimated amount consumed. METHODS: Between 2014 and 2015 samples of vomitus from seven dogs that ingested rodenticide baits containing BDF were submitted by veterinarians in New Zealand. All seven dogs had been given apomorphine by the veterinarian and vomited within 1 hour of ingesting the bait. Some or all of the bait particles were retrieved from each sample and were analysed for concentrations of BDF using HPLC. Based on estimations of the mass of bait consumed, the concentration of BDF stated on the product label, and the estimated mass of bait in the vomitus of each dog, the amount of BDF in the vomited bait was calculated as a percentage of the amount ingested. RESULTS: For five dogs an estimation of the mass of bait ingested was provided by the submitting veterinarian. For these dogs the estimated percentage of BDF in the bait retrieved from the vomitus was between 10-77%. All dogs were well after discharge but only one dog returned for further testing. This dog had a normal prothrombin time 3 days after ingestion. CONCLUSIONS AND CLINICAL RELEVANCE: The induction of emesis within 1 hour of ingestion can be a useful tool in reducing the exposure of dogs to a toxic dose of BDF. The BDF was not fully absorbed within 1 hour of ingestion suggesting that the early induction of emesis can remove bait containing BDF before it can be fully absorbed.

White snakeroot poisoning in goats: Variations in toxicity with different plant chemotypes.

Tremetone and possibly other benzofuran ketones are believed to be the toxic compounds in white snakeroot. However, disease has not been reproduced with purified toxins and the concentrations of the benzofuran ketones in white snakeroot populations that cause toxicosis have not been documented. The objectives of this study were to compare the toxicity of seven plant populations, better characterize the clinical and pathologic changes of poisoning, and correlate intoxication with benzofuran ketone content. Four of the seven white snakeroot collections were toxic at the dose and duration used in the study. Affected goats became exercise intolerant, had significant serum enzyme changes and histological lesions in the large appendicular muscles. The incidence and severity of poisoning was not correlated with total doses of tremetone or total benzofuran ketone concentrations suggesting they may not be closely involved in producing toxicity and the possible involvement of an unidentified toxin. The results also demonstrate that white snakeroot populations vary chemically and toxicologically.

Liver biopsy histopathology for diagnosis of Johne's disease in sheep.

Sheep with Johne's disease develop epithelioid macrophage microgranulomas, specific to Mycobacterium avium subsp. paratuberculosis (Map) infection, in the terminal ileum, mesenteric lymph nodes, and organs distant to the alimentary tract such as the liver. The objectives of this study were to determine whether liver pathology was present in ewes affected by Map and whether liver cores provide adequate tissue for this potential diagnostic marker. One hundred and twenty-six adult, low body condition ewes were euthanized, necropsied, and underwent simulated liver biopsy. Ileal lesions typical of Map were found in 60 ewes. Hepatic epithelioid microgranulomas were observed in all ewes with Type 3b (n = 40) and 82% (n = 11) with Type 3c ileal lesions. None were found in ewes unaffected by Map or with Type 1, 2, or 3a ileal lesions. Liver biopsy core samples provided adequate tissue for histopathology with a sensitivity and specificity of 96% (95% confidence interval [CI], 0.87-0.99) and 100% (95% CI, 0.95-1), respectively for detection of types 3b and 3c ileal lesions.

Bile duct lesions associated with turnip (Brassica rapa) photosensitization compared with those due to sporidesmin toxicosis in dairy cows.

Cattle grazing turnips or other brassica forage crops occasionally develop hepatogenous photosensitization. In New Zealand, cases of bovine photosensitization associated with such crops frequently occur during late summer and fall, and this coincides with the facial eczema (sporidesmin toxicosis) "season." Clinical chemistry findings in acute photosensitization cases associated with both brassica and facial eczema include marked serum elevations in γ-glutamyl transferase and glutamate dehydrogenase activities. Distinctive bile duct lesions of "subacute" turnip photosensitization in 2 cows, comprising microscopic cholangiectasis with concentric periductal fibrosis of small bile ducts, and a spectrum of changes from bile duct necrosis progressing to obliterative sclerosis are described. These bile duct lesions are compared with those in 3 cases of "subacute" facial eczema in adult cows, where medium-sized and larger ducts tend to be involved and bile duct hyperplasia and portal fibrosis are more prominent, often leading to bridging between neighboring portal triads.

The prevalence of disseminated Mycobacterium avium subsp. paratuberculosis infection in tissues of healthy ewes from a New Zealand farm with Johne's disease present.

AIM: To determine the prevalence of disseminated Mycobacterium avium subsp. paratuberculosis (Map) infection in healthy ewes in a flock with a history of clinical Johne's disease. METHODS: Twenty-four healthy ewes, from a large sheep and cattle farm with a history of clinical Johne's disease in the ewe flock, were randomly selected, euthanased, blood sampled, and examined at necropsy. BACTEC™ radiometric culture for Map was performed on samples of faeces, ileum, mesenteric lymph node, biceps femoris muscle and mononuclear cells in peripheral blood. Serum antibody ELISA tests were performed. Histological sections and Ziehl Neelsen (ZN) stains of impression smears of ileum and mesenteric lymph node were examined for pathological lesions characteristic of Johne's disease and acid fast organisms (AFO). Indirect quantification of Map was performed, using BACTEC radiometric growth indices measuring the time taken for the production of (14)CO(2.) RESULTS: No histological evidence of Johne's disease or AFO was found in the ileum and mesenteric lymph nodes. Twelve of the 24 ewes (50%) had Map cultured from the ileum (n=6) and/or mesenteric lymph nodes (n=8) while none had Map cultured from the faeces, biceps femoris muscle or blood mononuclear cells. One of the 12 Map culture positive ewes was serum ELISA positive. The culture growth rates in liquid medium suggest low numbers of Map were present in the tissues of the culture positive ewes. CONCLUSION: Fifty per cent of clinically healthy ewes exposed to Map within a Johne's infected flock were Map culture positive in the ileum and/or mesenteric lymph node(s), while the ELISA was positive in 8% of those animals (n=1). There was no faecal shedding of Map and no Map was cultured from skeletal muscle or from blood mononuclear cells suggesting that systemic Map infection, defined as positive culture of Map from skeletal muscle and/or blood, may be uncommon in healthy mixed age ewes without clinical Johne's disease. CLINICAL RELEVANCE: ELISA serology detected 1 of 12 ewes infected with Map whilst none were detected from faecal BACTEC radiometric culture, suggesting biosecurity measures used to control the spread of Map may be of limited use. Map was not cultured from blood mononuclear cells or skeletal muscle, indicating that meat from healthy ewes, from farms where Johne's disease is present, is an unlikely source of Map exposure for humans. Further research is warranted to establish the prevalence and dissemination of Map in tissues outside the alimentary tract of healthy ewes from farms throughout New Zealand where Map is present.

Pathology of Brucella ovis infection in red deer stags (Cervus elaphus).

AIM: To describe the pathology of the reproductive tract of red deer stags with active Brucella ovis infection and in stags in which B. ovis infection had resolved. METHODS: Twenty-three red deer stags of varying history were slaughtered and their epididymides and accessory sex glands examined grossly and by histopathology. At the time of slaughter five of the stags had an active B. ovis infection of 24-55 days duration following exposure to infected rams, 10 stags had been experimentally infected with B. ovis by intravenous inoculation 649 days previously and had developed an active infection but the bacterial infection had resolved at least 308 days prior to slaughter, and eight stags had not been exposed to B. ovis at any time. RESULTS: Of the five stags with an active infection, one had gross enlargement of the epididymides that could be detected by scrotal palpation. Histological lesions in all five stags included mild to severe, predominantly non-suppurative epididymitis, vesiculitis, prostatitis and ampullitis, with neutrophil exudation in associated glandular ducts. Additional lesions in the epididymides were spermatic granulomas and epithelial hyperplasia with intra-epithelial cyst formation. Of the 10 stags in which the bacterial infection had resolved, two had gross enlargement of the epididymides. The histological lesions were similar to those in stags with active infection but were generally milder, with increased periductal scar tissue in the epididymides. The lesions seen in stags resembled those seen in rams with B. ovis infection but they were usually less florid and had fewer plasma cells. No gross abnormalities or histopathological lesions were detected in the non-infected stags. CONCLUSIONS: Only a small percentage of red deer stags infected with B. ovis develop lesions of epididymitis that can be detected by scrotal palpation. Gross and histological lesions of the genital tract of stags associated with B. ovis infection are similar to the lesions seen in rams. Lesions in stags persist for >300 days after the bacterial infection has resolved. CLINICAL RELEVANCE: Brucella ovis infection should be considered when there are gross lesions of epididymitis or histological evidence of inflammation in the epididymides or accessory sex glands of red deer stags. Retrospective diagnosis of B. ovis in stags could be achieved by histological examination of the reproductive organs.

Distal aortic aneurysm presumed to be secondary to an infected umbilical artery in a foal.

CASE HISTORY: A 3-month-old female Warmblood foal was presented after displaying signs of colic with pyrexia for 5 days. CLINICAL AND PATHOLOGICAL FINDINGS: The foal continued to show signs of colic, frequently passed urine, and was pyrexic with an elevated white blood cell count. The umbilical stalk was thickened but there was no evidence of purulent material. Exploratory laparotomy revealed an enlarged left umbilical artery remnant tightly adhered to the bladder wall. The left umbilical artery continued to an aneurysm involving the distal aorta. The foal was subject to euthanasia and post-mortem examination confirmed a spherical aortic aneurysm, in the dorsal midline caudal to the kidneys that contained a large thrombus. Histopathological examination revealed inflammation and necrosis of the tunica intima and tunica media of the left umbilical artery with suppuration and bacterial colonies evident in the periarterial tissues. DIAGNOSIS: Infected aortic aneurysm presumably caused by an umbilical artery infection. CLINICAL RELEVANCE: A previously undetected umbilical infection appears to have resulted in an unusual delayed complication causing signs of colic in a foal. Veterinarians should be aware of this condition, and the possibility that it may be a cause of signs of colic in foals. Diagnosis based on ultrasonography should be possible, but may require sedation, visceral analgesia and careful examination.

Potential involvement of Neospora caninum in naturally occurring ovine abortions in New Zealand.

Neospora caninum is an obligate intracellular parasite and is recognised as the leading cause of bovine abortion worldwide. Natural infection with N. caninum has been described in sheep but it has generally not been regarded as a significant cause of abortion. Recently, there have been several New Zealand cases of foetal abortions where N. caninum was detected which strongly suggested the involvement of Neospora in these abortions. However, there is minimal information about the prevalence of N. caninum infection naturally occurring in New Zealand sheep flocks and particularly its impact on reproduction success. Thus, this present study provides preliminary data on the role that Neospora is playing in ovine reproductive failure by establishing the prevalence of N. caninum antibodies and DNA in ewe blood and foetal material present in 21 New Zealand sheep farms with ongoing unexplained abortion problems and 10 farms with consistently high fertility levels. The results of this study demonstrated an overall seroprevalence of 1.4% which varied between Aborting/non-pregnant (1.8%), age-matched pregnant controls (0.6%) and high fertility (2.1%) ewes. However, despite the variation observed, there was no statistical difference between the three groups. In addition, Neospora DNA was detected by PCR in 13% of submitted foetal brains and in ewe blood from aborting/non-pregnant (6.9%), age-matched pregnant controls (3.6%) and high fertility pregnant (2.1%) ewes. When the PCR results were considered with the IFAT and IDEXX ELISA results, there was no correlation between serology positive and PCR positive blood samples. Taken together, these results reveal that reliance on ELISA-based serology or PCR alone may underestimate the involvement of Neospora. Furthermore, determining the involvement of Neospora appears to require a multi-facetted approach where diagnostic methods and serological cut-off values may need to be adjusted as further information about the effect of natural infections with N. caninum in the ovine host is elucidated.

Detection of Mycobacterium avium subsp. paratuberculosis in skeletal muscle and blood of ewes from a sheep farm in New Zealand.

AIM: To determine whether viable Mycobacterium avium subsp. paratuberculosis (Map) is present in skeletal muscle and blood in ewes with and without Johne's disease confirmed histologically. METHODS: A total of 51 mixed-aged ewes in poor body condition from a farm with a history of clinical Johne's disease were culled and examined at necropsy. BACTEC radiometric culture was performed on samples of skeletal muscle from the biceps femoris, mononuclear cells in peripheral blood (hereafter referred to as blood), and ileum. Histological sections and Ziehl-Neelsen (ZN)-stained impression smears of terminal ileum and mesenteric lymph nodes were examined. Ewes were defined as having confirmed Johne's disease if there was histopathological evidence typical of the disease within the ileum and adjacent lymph nodes. RESULTS: Eighteen of 21 (86%) ewes with confirmed clinical Johne's disease were culture-positive for Map from sites peripheral to the alimentary tract, comprising 15 from skeletal muscle and 13 from blood. Five of 30 (17%) ewes that did not have Johne's disease were culture-positive, with four from skeletal muscle and one from blood. The likelihood that ewes with confirmed Johne's disease had systemic Map infection compared with ewes without was determined as OR=30 (95% CI=6.3-142.0; p<0.001). CONCLUSION: The prevalence of Map infection of skeletal muscle and blood in ewes with confirmed Johne's disease was 71% and 62% respectively, and in unaffected ewes was 13% for muscle and 3% for blood. CLINICAL RELEVANCE: Skeletal muscle and blood are potential sources of exposure of humans to Map, and the risk appears higher from sheep with Johne's disease.

Photosensitisation, crystal-associated cholangiohepatopathy, and acute renal tubular necrosis in calves following ingestion of Phytolacca octandra (inkweed).

CASE HISTORY: In March 2006, an outbreak of photosensitivity affecting 6-8-month-old Friesian heifer calves on a farm in the Rangitikei district of New Zealand was investigated. The calves were grazing wheat stubble paddocks that also had a variety of weeds, especially Phytolacca octandra (inkweed). They also had access to pond water that contained potentially toxic concentrations of the cyanobacteria (blue-green algae) Microcystis and Planktothrix spp. CLINICAL FINDINGS: Initially, affected animals showed acute irritation, agitation, reluctance to walk, recumbency in some animals, hyperaemia of unpigmented skin and jaundice. Serum chemistry revealed elevated liver enzyme activities and azotaemia. Later in the outbreak, exudative dermatitis with formation of crusts on unpigmented skin, dehydration and inappetence were notable signs, as well as occasional diarrhoea. PATHOLOGICAL AND TOXICOLOGICAL FINDINGS: Post-mortem examinations following euthanasia of four severely affected calves and a fifth animal that died naturally revealed livers that were grossly orange brown and mildly enlarged, and kidneys that were enlarged and pale brown. Microscopic lesions in the liver were mild; however, small birefringent crystals in the bile ducts were noted in the first two cases. In contrast, renal lesions were moderate to severe, and included prominent segmental tubular necrosis, granular and cellular casts, and mild interstitial non-suppurative inflammation. In the kidneys of animals examined later in the outbreak, there was early interstitial fibrosis as well as tubular regeneration, with numerous hyaline casts in the renal medulla. Inkweed plants had been heavily browsed, and recognisable portions of the plant were found in the gastrointestinal tracts of affected calves. Chemical analysis of inkweed material revealed triterpene saponins. No known hepatotoxic or nephrotoxic plants were identified in the paddocks. The hepatic lesions were not consistent with published descriptions of cyanobacterial toxicity. Sporidesmin toxicity was ruled out. DIAGNOSIS: Hepatogenous photosensitivity, crystal-associated cholangiohepatopathy, toxic acute renal tubular necrosis, associated with the ingestion of P. octandra, and possibly complicated by cyanobacteria in the water. CLINICAL RELEVANCE: Crystal-associated cholangiohepatopathy with photosensitivity in cattle is rare, and has only been reported in steroidal saponin-containing Brachiaria decumbens poisoning, in Brazil. The consistent pattern of toxic acute renal tubular necrosis was similar to that caused by the ingestion of Quercus, Amaranthus or Lantana spp. A combination of toxicities was conceivable but circumstantial evidence strongly implicated P. octandra. Further toxicological investigation of this plant is warranted before it can be listed as a known nephrotoxin of cattle.

Photodynamic chlorophyll a metabolites, including phytoporphyrin (phylloerythrin), in the blood of photosensitive livestock: overview and measurement.

AIM: To validate a spectrofluorometric method for measuring chlorophyll a metabolites, specifically phytoporphyrin (= phylloerythrin), as well as the chlorins, pheophorbide a and pyropheophorbide a, in the blood of photosensitive cattle and sheep. METHODS: Standard methanolic solutions of pheophorbide a (25 microM), pyropheophorbide a (25 microM), and phytoporphyrin (<3.7 microM) were prepared. Serum and plasma samples were obtained from cattle (n=5), sheep (n=3), and one alpaca, with clinical facial eczema (i.e. photosensitive), as well as from clinically normal (n=2 of each) adult cows, recently weaned calves, and sheep (controls). Standard solutions of the three metabolites were characterised using high-performance liquid chromatography (HPLC), with mass spectrometry, in conjunction with absorption and emission spectral data, and were compared with sera from photosensitive animals. In the latter, phytoporphyrin was the only metabolite detected. Calibration curves were prepared by adding different ratios of methanol and standard solutions of phytoporphyrin in methanol to diluted serum from control animals. Peak areas of fluorescence spectra were determined in samples from photosensitive animals. RESULTS: Pheophorbide a and pyropheophorbide a produced typical chlorin spectra, and had excitation/emission maxima of 408/669 nm and 409/669 nm, respectively. Phytoporphyrin showed a typical porphyrin fluorescence spectrum, with excitation/ emission maxima of 425/644 nm. Pyropheophorbide a and phytoporphyrin had very similar chromatographic retention times, the same chemical formula and same mass, but were distinguishable by differences in their absorption spectra. In sera from photosensitive animals, the fluorescence emission at 644 nm was shown to arise solely from phytoporphyrin and not from any other chlorophyll a metabolites. Calibration curves using sera and plasma from control animals gave reliable measurements of phytoporphyrin in the range 0.4-6 microM. The sera of facial eczema-affected cattle and sheep had concentrations of phytoporphyrin ranging from 0.4 to 1.8 and 0.9 to 2.8 microM, respectively. Haemolysed serum samples were not suitable for determination of phytoporphyrin with this method. CONCLUSIONS AND CLINICAL RELEVANCE: A spectrofluorometric method for the quantification of phytoporphyrin in the blood of photosensitive animals has been validated, and can be applied to the measurement of other chlorophyll a metabolites in blood. This will be a useful tool in the further investigation of the cause and pathogenesis of idiopathic photosensitivities of farm animals.