Hematology and plasma chemistry values in free-ranging Humboldt penguins (Spheniscus humboldti) in Chile

To establish baseline hematologic and plasma biochemistry values in free-ranging Humboldt penguins (Spheniscus humboldti), heparinized blood samples were collected from 51 apparently healthy, adult Humboldt penguins residing at two colonies off the Chilean coast. Thirty samples were collected in April, 1992, from penguins inhabiting the Ex-islote de los Pájaros Niños in Algarrobo, Chile. In September, 1992, 21 samples were collected from birds inhabiting Isla de Cachagua, Chile. Hematologic values measured include packed cell volume, leucocyte count, leucocyte differential, and the presence of blood parasites. Plasma biochemistry values measured include glucose, blood urea nitrogen, creatinine, uric acid, calcium, inorganic phosphorous, sodium, potassium, chloride, total protein, albumin, globulin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total bilirubin, and creatine kinase. Only the mean values for chloride and for the number of eosinophils differed significantly between the two sample groups. No blood parasites were seen. © 1995 Wiley-Liss, Inc.     

Avian disease at the Salton Sea

A review of existing records and the scientific literature was conducted for occurrences of avian diseases affecting free-ranging avifauna within the Salton Sea ecosystem. The period for evaluation was 1907 through 1999. Records of the U.S. Department of Agriculture, Bureau of Biological Survey and the scientific literature were the data sources for the period of 1907–1939. The narrative reports of the U.S. Fish and Wildlife Service's Sonny Bono National Wildlife Refuge Complex and the epizootic database of the U.S. Geological Survey's National Wildlife Health Center were the primary data sources for the remainder of the evaluation. The pattern of avian disease at the Salton Sea has changed greatly over time. Relative to past decades, there was a greater frequency of major outbreaks of avian disease at the Salton Sea during the 1990s than in previous decades, a greater variety of disease agents causing epizootics, and apparent chronic increases in the attrition of birds from disease. Avian mortality was high for about a decade beginning during the mid-1920s, diminished substantially by the 1940s and was at low to moderate levels until the 1990s when it reached the highest levels reported. Avian botulism (Clostridium botulinum type C) was the only major cause of avian disease until 1979 when the first major epizootic of avian cholera (Pasteurella multocidia) was documented. Waterfowl and shorebirds were the primary species affected by avian botulism. A broader spectrum of species have been killed by avian cholera but waterfowl have suffered the greatest losses. Avian cholera reappeared in 1983 and has joined avian botulism as a recurring cause of avian mortality. In 1989, avian salmonellosis (Salmonella typhimurium) was first diagnosed as a major cause of avian disease within the Salton Sea ecosystem and has since reappeared several times, primarily among cattle egrets (Bubulcus ibis). The largest loss from a single epizootic occurred in 1992, when an estimated 155000 birds, primarily eared grebes (Podiceps nigricollis), died from an undiagnosed cause. Reoccurrences of that unknown malady have continued to kill substantial numbers of eared grebes throughout the 1990s. The first major epizootic of type C avian botulism in fish-eating birds occurred in 1996 and killed large numbers of pelicans (Pelecanus occidentalis & P. erythrorhynchos). Avian botulism has remained as a major annual cause of disease in pelicans. In contrast, the chronic on-Sea occurrence of avian botulism in waterfowl and shorebirds of previous decades was seldom seen during the 1990s. Newcastle disease became the first viral disease to cause major bird losses at the Salton Sea when it appeared in the Mullet Island cormorant (Phalacrocorax auritus) breeding colony during 1997 and again during 1998.     

Environmental Factors Influencing the Prevalence of a Clostridium botulinum Type C/D Mosaic Strain in Nonpermanent Mediterranean Wetlands

Between 1978 and 2008, 13 avian botulism outbreaks were recorded in the wetlands of Mancha Húmeda (central Spain). These outbreaks caused the deaths of around 20,000 birds from over 50 species, including globally endangered white-headed ducks (Oxyura leucoceophala). Here, a significant association was found between the number of dead birds recorded in each botulism outbreak and the mean temperature in July (always >26°C). The presence of Clostridium botulinum type C/D in wetland sediments was detected by real-time PCR (quantitative PCR [qPCR]) in 5.8% of 207 samples collected between 2005 and 2008. Low concentrations of Cl⁻ and high organic matter content in sediments were significantly associated with the presence of C. botulinum. Seventy-five digestive tracts of birds found dead during botulism outbreaks were analyzed; C. botulinum was present in 38.7% of them. The prevalence of C. botulinum was 18.2% (n = 22 pools) in aquatic invertebrates (Chironomidae and Corixidae families) and 33.3% (n = 18 pools) in necrophagous invertebrates (Sarcophagidae and Calliphoridae families), including two pools of adult necrophagous flies collected around bird carcasses. The presence of the bacteria in the adult fly form opens up new perspectives in the epidemiology of avian botulism, since these flies may be transporting C. botulinum from one carcass to another.     

Immunization of ducks for type C botulism

A single subcutaneous immunization with a vaccine used for protecting ranch mink (Mustela vison) against type C botulism reduced morbidity and mortality in mallard (Anas platyrhynchos) and northern pintail (Anas acuta) ducks challenged with approximately 4.5 x 10(4) and 2.25 x 10(4) mouse lethal doses (MLD50), respectively, of botulinum toxin at 10 and 15 days post-immunization (pi). There was no significant protection at 5 days pi. Protection persisted in mallards for 90 days pi. To simulate use of vaccine as a part of treatment of sick birds in the field, mallards were exposed to toxin and, when clinical signs were evident, each bird was treated by intraperitoneal injection of type C botulinum antitoxin and one-half of the birds were immunized. Immunization had no significant effect on recovery from intoxication. At 10 days posttreatment, all birds were challenged with toxin. Clinical signs and mortality were significantly less frequent among immunized birds than among non-immunized birds after the second exposure. Immunization might be useful as part of the treatment regimen in botulism outbreaks.     

Hematology and plasma biochemistry reference range values for free-ranging desert tortoises in Arizona

Baseline values and ranges for 10 hematologic and 32 plasma chemistry parameters were analyzed for 36 free-ranging Sonoran desert tortoises (Gopherus agassizzi) collected in Yavapai and La Paz Counties (Arizona, USA) from 1990 to 1995. Tortoises were radio tagged from 1990 to 1994, and attempts were made to recapture them three times a year. Tortoises were weighed, measured, and chemically immobilized to collect blood for hematology and blood chemistry assessments. Tortoise biochemistry differed (P < 0.01) between sites and sexes and among seasons and years. Normal reference ranges for hematologic and plasma biochemistry parameters were determined. Seasonal and annual differences in hematology and blood chemistry were related to rainfall patterns, forage availability, and physiological condition.     

Hematology, plasma chemistry, and bacteriology of wild Tundra Swans (Cygnus columbianus) in Alaska

Blood and cloacal swabs were collected from 100 (66 female, 34 male) wild Tundra Swans (Cygnus columbianus) molting in northwestern Alaska, USA, 25-28 July 2008, to establish hematologic and serum chemistry reference values and to isolate enteric Salmonella spp. and Escherichia coli O157:H7. Plasma biochemistry and hematology values did not vary significantly by sex or age. Tundra swans had high levels of creatine kinase, lactate dehydrogenase, amylase, and alkaline phosphatase compared with some other avian species (values were up to 7 times greater), possibly indicating capture myopathy. However, concentrations were much lower (up to 8 times lower) than in other waterfowl exposed to similar or more intensive capture methods. White blood cell count and hematocrit values were similar to other waterfowl species, and enteric Salmonella spp. and E. coli O157:H7 were not present among birds sampled. Our data provide the first biochemical, hematologic, and bacteriologic reference values for wild Tundra Swans.     

Hematologic, protein electrophoresis, biochemistry, and cholinesterase values of free-living black stork nestlings (Ciconia nigra)

Hematologic, protein electrophoresis, serum biochemistry, and cholinesterase values were determined in 36 free-living black stork nestlings (Ciconia nigra) between 25 and 53 days of age in order to establish normal reference values for this population. The following values were evaluated: white blood cell counts, red blood cell counts, packed cell volume, hemoglobin, heterophils, lymphocytes, monocytes, eosinophils, prealbumin, albumin, alpha-globulin, beta-globulin, gamma-globulin, total protein, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, creatine kinase, calcium, phosphorus, iron, cholesterol, glucose, triglycerides, uric acid, urea, creatinine, total solids, bile acids, and butyrylcholinesterase. Sex-dependent differences were observed in hemoglobin, prealbumin, albumin, gamma-globulin, total protein, alkaline phosphatase, and triglycerides. Packed cell volume, butyrylcholinesterase, aspartate aminotransferase, creatine kinase, and creatinine increased with age, whereas albumin, mean cell volume, calcium, phosphorus, cholesterol, and total solids decreased with age. These hematologic and serum biochemistry values can be used as reference ranges in free-living black stork nestlings.     

Brain acetylcholinesterase activity in botulism-intoxicated mallards

Brain acetylcholinesterase (AChE) activity in captive-reared mallards (Anas platyrhynchos) that died of botulism was compared with euthanized controls. AChE levels for both groups were within the range reported for normal mallards, and there was no significant difference in mean AChE activity between birds that ingested botulism toxin and died and those that did not.     

A relationship between avian carcasses and living invertebrates in the epizootiology of avian botulism.

A survey of the sources of Clostridium botulinum type C toxin possibly utilized as food by aquatic birds in an epizootic area of avian botulism in northern Utah showed that living aquatic and terrestrial invertebrates normally found in close association with dead, decomposing birds commonly carried the toxin. Of 461 samples associated with 21 species of avian carcasses, 198 were toxin-positive. Invertebrate species not normally scavengers of vertebrate tissues were less commonly and less highly toxic, particularly when captured 30 cm or more from a carcass; six of 237 samples of such aquatic invertebrates low-level toxin. Of the species tested, blow fly larvae (Calliphoridae) were the most consistently and highly toxic, although others, particularly adult and larval stages of several species of beetles (Coleoptera), contained toxin at levels probably significant in the epizootiology of the disease. An estimated 0.05 to 0.25 g of the most toxic fly larvae or 15 g of the most toxic beetles tested carried a mediam lethal dose for an adult mallard duck. Examination of stomach contents of aquatic birds dead of botulism showed that some had consumed invertebrates.     

Biodiversity of Clostridium botulinum type E associated with a large outbreak of botulism in wildlife from Lake Erie and Lake Ontario

The genetic relatedness of Clostridium botulinum type E isolates associated with an outbreak of wildlife botulism was studied using random amplification of polymorphic DNA (RAPD). Specimens were collected from November 2000 to December 2008 during a large outbreak of botulism affecting birds and fish living in and around Lake Erie and Lake Ontario. In our present study, a total of 355 wildlife samples were tested for the presence of botulinum toxin and/or organisms. Type E botulinum toxin was detected in 110 samples from birds, 12 samples from fish, and 2 samples from mammals. Sediment samples from Lake Erie were also examined for the presence of C. botulinum. Fifteen of 17 sediment samples were positive for the presence of C. botulinum type E. Eighty-one C. botulinum isolates were obtained from plants, animals, and sediments; of these isolates, 44 C. botulinum isolates produced type E toxin, as determined by mouse bioassay, while the remaining 37 isolates were not toxic for mice. All toxin-producing isolates were typed by RAPD; that analysis showed 12 different RAPD types and multiple subtypes. Our study thus demonstrates that multiple genetically distinct strains of C. botulinum were involved in the present outbreak of wildlife botulism. We found that C. botulinum type E is present in the sediments of Lake Erie and that a large range of bird and fish species is affected.     

Hematology and serum biochemistry of the brush-tailed rock-wallaby (Petrogale penicillata)

In Australia the brush-tailed rock-wallaby (Petrogale penicillata) is the subject of a national recovery plan, and several sites have been selected for reintroductions. Condition of wild populations and individual animals can be monitored using hematologic and serum biochemistry analytes, and hematologic variables have been correlated with postrelease survival in other species. Prior to such monitoring, reference values for blood variables are required, but these data have not been available for the brush-tailed rock-wallaby. During four trapping periods from November 2004 to August 2005, 116 blood samples were collected from 44 brush-tailed rock-wallabies in a wild colony in southeast Queensland. Some variables varied with sex, age, method of restraint, lactation demands, and trapping period. After partitioning, when required, reference ranges for hematology and serum biochemistry variables were established. This study provides the most comprehensive serum biochemistry reference range for any macropodid marsupial yet published.     

PCR-based molecular diagnosis of botulism (types C and D) outbreaks in aquatic birds

Clostridium botulinum is a strictly anaerobic spore forming bacterium found in soil and sediments, producing botulinum neurotoxins. Group III strains of this organism are only able to produce neurotoxin types C, mosaic C/D, D/C, and D, which are associated with bird botulism. The threats and outbreak cycle of bird botulism are enhanced in the aquatic environment via a food web-environment-avian interface in response to global climatic changes. The aim of this review was to describe and discuss the development of PCR-based markers, diagnostic assays, and applications with special emphasis to botulism detection in aquatic birds. We employed a text-mining approach for collection of current diagnostic information to bird botulism in aquatic environments. Using the PubMed search engine, we have comprehensively collected relevant information from 124 articles and then summated for the purpose of this review. Genes coding for botulinum neurotoxin (bont/C, bont/C/D, bont/D/C, and bont/D), nontoxic components (ha70, ha17, ha33, and ntnh), and flagellin (fliC) were the molecular markers most commonly found for the genotyping of group III strains from environmental samples. “GeneDisc” real-time PCR system was a robust and reliable diagnostic technique for discrimination of neurotoxin types in the large sampled areas. PCR-based assays were the most perceptive and widely established for detection of bird botulism, even if several biochemical and molecular diagnostic techniques have been available. Thus, timely and accurate identification of its mortality is needed to provide a biosecurity, disease control management, and conservation of aquatic birds. PCR-based diagnosis is a promising alternative to the mouse bioassay. Hence, the present paper makes an attempt to review the PCR-based detection assays including primers, specificity, sensitivity, and detection limit and explore their potential applications in wildlife microbiology.     

Toxicity of Clostridium botulinum type E neurotoxin to Great Lakes fish: implications for avian botulism

Since 1999, large-scale mortalities of fish-eating birds have been observed on the Great Lakes, and more specifically on Lake Erie. Type E botulism has been established as the primary cause of death. The mechanism of type E botulism exposure in fish-eating birds is unclear. Given that these birds are thought to eat live fish exclusively, it seems likely that their prey play a key role in the process, but the role of fish as potential transport vectors of botulinum neurotoxin type E (BoNT/E) to birds has not been adequately investigated. Between June 2003 and April 2004 a methodological model for exposing fish to Clostridium botulinum was developed and used to compare the sensitivity of rainbow trout (Oncorhynchus mykiss), round goby (Neogobius melanostomas), walleye (Stizostedion vitreum), and yellow perch (Perca flavescens) to four doses (0, 800, 1,500, and 4,000 Mouse Lethal Doses) of Clostridium botulinum type E neurotoxin. Each fish species expressed unique changes in both behavior and skin pigmentation prior to death. Yellow perch survived significantly longer (P < 0.05) than the three other species at all toxin treatments. Results of this study suggest that live fish can represent a significant vector for transfer of BoNT/E to birds.     

Tick infestations of birds in coastal Georgia and Alabama

Mist-netted birds were examined for ticks on Jekyll Island, Glynn Co., Georgia (32 bird species) in 1996-1998, and at Fort Morgan, Baldwin Co., Alabama (36 species) in 1998 during fall migration. Sixty-two (14.7%) of 423 birds from Jekyll Island and 22 (13.3%) of 165 birds from Fort Morgan were infested with ticks. The mean number of ticks per infested bird was 2.0 on Jekyll Island and 6.3 at Fort Morgan. Ten species of birds were infested with ticks on Jekyl1 Island where 87% of all ticks were recovered from 3 species: the common yellowthroat (Geothlypis trichas), gray catbird (Dumetella carolinensis), and northern waterthrush (Seiurus noveboracensis). Eight species of birds were infested with ticks at Fort Morgan where 83% of all ticks were recovered from 3 species: the brown thrasher (Toxostoma rufum), swamp sparrow (Melospiza georgiana), and common yellowthroat. Six species of ticks (Amblyomma americanum, Amblyomma maculatum, Haemaphysalis leporispalustris, Ixodes brunneus, Ixodes minor, and Ixodes scapularis) were recovered from the Georgia birds, whereas 3 species (A. maculatum, H. leporispalustris, and Ixodes dentatus) were recovered from the Alabama birds. Attempts to isolate Borrelia burgdorferi sensu stricto, the etiologic agent of Lyme borreliosis, from Ixodes spp. ticks recovered from birds were unsuccessful.     

Hematology and serum chemistry of the island fox on Santa Cruz Island

Serum and hematologic biochemistry values for island foxes (Urocyon littoralis) on Santa Cruz Island (California, USA) in April (wet season) and September (dry season) 1998 were evaluated. Serum chemistry of island foxes generally varied seasonally; 10 (40%) of the 25 serum characteristics were higher in the wet season, and three (12%) of the 25 serum characteristics were higher in the dry season. No hematologic parameters varied between seasons, although some measures varied between sexes. Blood analytes also varied with age; fox pups had higher values than adults for one hematologic and four serum parameters, whereas adult foxes had higher values for five hematologic characteristics. The information on blood chemistry provides baseline data useful in the monitoring of this threatened insular endemic carnivore.     

Outbreaks of type C botulism in waterfowl in Japan

Four outbreaks of botulism in waterfowl were encountered over a five-year period of 1973 to 1977 in Japan. In all the outbreaks toxin was detected from all 12 sera, twenty-three of 24 gizzard contents from diseased or dead birds and one of three maggots. It was neutralized with Clostridium botulinum type C antitoxin serum, regardless of its origin. By using CO2 gas jet method, C. botulinum was isolated from four of 11 gizzards from diseased birds, five of 7 ones from dead birds, one of one maggot and one of one sludge sample, that is, eleven of 20 specimens in total. All 20 strains were identical with C. botulinum type C in biological properties. Most of the isolates showed a toxin titer ranging from 1,000 to 200,000 LD50 for mice. Four of them were identified as type C by mouse neutralization tests with antitoxin sera. The toxic suspensions of a strain 1-15 were administered orally to Chinese spot-billed ducks, which died when more than 200,000 LD50 mouse toxin was administered. Environmental conditions for occurrences of waterfowl botulism were discussed.     

Protective effects of vitamins (C and E) and melatonin co-administration on hematological and hepatic functions and oxidative stress in alloxan-induced diabetic rats

The present study aimed to investigate the potential effects of vitamins (C and E)/melatonin co-administration on the hematologic and hepatic functions and oxidative stress in alloxan-induced diabetic rats. The intraperitoneal injection of alloxan (120 mg/kg b.w. for 2 days) induced a significant increase of blood glucose levels (hyperglycemia) associated with serious hematologic disorders (P?<?0.01) evidenced by the decrease in the levels of red blood cell count (RBC) (?18 %), hematocrit (Ht) (?18 %), hemoglobin content (Hb) (?36 %), mean corpuscular hemoglobin (MCH) (?17 %), and mean corpuscular hemoglobin concentration (MCHC) (?16 %). The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and the plasmatic levels of total cholesterol and triglyceride contents of diabetic rats were, however, noted to undergo significant increases by 42 % (P?<?0.01), 134 % (P?<?0.001), 27.5 % (P?<?0.01), 147 % (P?<?0.001), and 67 % (P?<?0.01), respectively, as compared to the control animals. Furthermore, a significant increase in malondialdehyde (MDA) content and a significant decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities were observed in the plasma and hepatic tissues of diabetic rats when compared to the controls. Interestingly, the treatment with vitamins (C, E) in combination with melatonin was noted to reduce the plasma levels of glucose, lower the MDA levels, and restore the hematologic parameters and biochemical and antioxidant levels of diabetic rats back to normal values, alleviating diabetes metabolic disorders in rats.     

Effects of botulism on ducks drinking saline water

Mallard (Anas platyrhynchos) ducklings (2 wk old) were given water from natural saline wetlands or fresh water as drinking water for 1 or 2 wk prior to, and after, receiving material containing Clostridium botulinum type C toxin. Water with conductivity ranging from 3,460 to 6,690 mu mhos/cm had no detectable effect on the occurrence or severity of clinical signs of botulism. Ducks drinking water with conductivity of 7,130 mu mhos/cm for 1 wk prior to receiving toxin had more severe clinical signs and greater mortality than did birds drinking fresh water. Ducks given the same water for 2 wk prior to receiving toxin did not differ from the controls in response to toxin. Fewer ducks in groups drinking the most saline water tested (conductivity = 13,500 mu mhos/cm) had clinical signs of botulism than in groups drinking fresh water.     

肉毒中毒30例护理体会

目的：探讨肉毒中毒患者救治的护理措施和效果。方法：选择我院自2000年1月～2010年12月收治入院的30例临床确诊为肉毒中毒的患者,男12例,女18例,年龄11～72岁。在救治过程中,除了及早应用肉毒抗毒素外,应做好以下护理工作：护理评估、心理护理、饮食护理、呼吸护理及褥疮护理。结果：30例肉毒毒素中毒患者中,1例因病情危重、肺部感染、呼吸衰竭死亡外,其余均在1～4周痊愈出院。结论：对于肉毒毒素中毒患者,除了及早应用肉毒抗毒素外,护理工作对患者的救治起着重要的作用。     

Efficacy of a type C botulism vaccine in green-winged teal

We tested the efficacy of a single dose of Botumink toxoid for protecting wild green-winged teal (Anas crecca) during botulism epizootics caused by Clostridium botulinum type C. We challenged control and immunized ducks with four different doses of type C botulinum toxin to determine the LD50 for this species and to evaluate vaccine protection. Fewer immunized ducks were affected with botulism than control ducks, indicating that a single dose of Botumink toxoid could increase the survival of ducks during epizootics. However, the frequency of immunized ducks with signs of botulism increased with the challenge dose of botulinum toxin. Even at doses of botulinum toxin approximately 2 to 4 green-winged teal LD50, about 50% of the immunized ducks were affected. We believe an improved vaccine or a better delivery system is required to justify immunization of wild birds for experimental survival studies.