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Prenatal passive transfer of Mycobacterium tuberculosis antibodies in Asian elephant (Elephas maximus) calves
Abstract
Asian elephant (Elephas maximus) dams and their newborn calves were tested for Mycobacterium tuberculosis antibodies in serum. Blood was drawn from dams prior to calving and from calves on their day of birth. All six calves born to tuberculosis-reactive dams were also tuberculosis reactive, suggesting prenatal passive placental transfer of tuberculosis antibodies. In contrast, all three calves born to tuberculosis-nonreactive dams lacked detectable tuberculosis antibodies in pre-suckling or day-of-birth blood samples. Of the living tuberculosis-reactive calves observed from 1 to 11 yr of age, none exhibited clinical signs of tuberculosis infection or became tuberculosis culture positive. This is the first report of prenatal passive placental transfer of tuberculosis antibodies in elephants and demonstrates that detectible tuberculosis antibodies in newborn elephant calves should not be assumed to correlate with clinical tuberculosis.
... In this study, calves at birth had levels of anti-tetanus and anti-rabies antibodies equal to or greater than their dams (who had been vaccinated for both rabies and tetanus during pregnancy) before the intake of colostrum. McGee et al. (2014) had similar findings in their study of tuberculosis-positive dams and calves, where they found that calves born to tuberculosis-positive Asian elephant dams had detectable anti-tuberculosis antibodies in day-of-birth or presuckling samples compared to samples from calves who were born from tuberculosis-negative mothers, which had no detectable antibodies (McGee et al. 2014). ...
... In this study, calves at birth had levels of anti-tetanus and anti-rabies antibodies equal to or greater than their dams (who had been vaccinated for both rabies and tetanus during pregnancy) before the intake of colostrum. McGee et al. (2014) had similar findings in their study of tuberculosis-positive dams and calves, where they found that calves born to tuberculosis-positive Asian elephant dams had detectable anti-tuberculosis antibodies in day-of-birth or presuckling samples compared to samples from calves who were born from tuberculosis-negative mothers, which had no detectable antibodies (McGee et al. 2014). ...
Elephant immunology is an emerging field of study. While we still have much to learn about how the elephant immune system differs from the immune systems of other animals and what advantages or disadvantages these differences may provide, our knowledge of the elephant immune system continues to increase as we work to understand disease mechanisms that threaten these incredible animals. Elephant endotheliotropic herpesvirus (EEHV) remains one of the most serious threats to the health and survival of Asian elephants. Immune responses to this virus in elephants are under active investigation in an attempt to understand why some elephants are able to recover, while others rapidly die of progressive disease. The elephant immune responses to EEHV, and other infections in elephants, are a focus of this chapter, which reviews our current understanding of elephant immunology and points to gaps in our knowledge.
... 18 In African elephant calves, 6-21 hr may elapse before nursing. 9,41 The long interval between birth and nursing may reflect the fact that, unlike hoofstock, elephant calves are immunocompetent at birth 38,48 and thus do not have immediate need for colostrum. Additionally, glucose may not be urgently needed in newborn elephants due to adequate stores acquired prenatally. ...
Sixteen years of medical records documenting 19 births within a herd of Asian elephants ( Elephas maximus) at a private facility in the southeastern United States were reviewed. Of the 19 calves, 11 were normal at birth, requiring no additional veterinary care, and eight were abnormal, requiring veterinary care immediately or within the first week of birth. Descriptive statistics were used to evaluate morphometrics, vital signs, and behavioral milestones in newborn calves both normal and abnormal. Blood work and urinalysis results from all calves were compared to values for adult elephants. Medical management of abnormal calves is described. All calves had faster heart rates and respiratory rates than did adult elephants, but rectal temperatures were the same. Calves were precocious with regard to sitting and standing but could be very slow to nurse. The most-common medical conditions of newborn calves were umbilical abnormalities and problems associated with nursing. Two calves required cardiopulmonary resuscitation after birth but made full recoveries. Some conditions were not apparent at birth but were recognized a few hours or days later. Following veterinary intervention, six of the eight calves made full recoveries, suggesting that early identification and treatment of problems can greatly decrease mortality. This is the first report of multiple veterinary and behavioral parameters in normal and abnormal neonatal Asian elephants from a facility with a calf survival rate above 90%. This information may be helpful to other elephant-holding facilities in providing care to their newborn elephant calves.
An 18-yr-old female Asian elephant (Elephas maximus) gave birth to a 120-kg female calf following 22 mo of gestation. Immediately after parturition, the cow became agitated and aggressive towards the calf. Before the keepers were able to safely intervene and remove the calf, the cow stepped on the calf's head and right front leg. Within 30 min, the cow calmed down, allowing the calf's safe reintroduction under close keeper supervision and control. The cow had a retained placenta, poor mammary development, and low milk production. The calf's injuries, in combination with the cow's low milk production, impeded the calf's ability to nurse and gain weight. Within 10 days, the calf lost 10% of its weight. Serum protein electrophoresis indicated failure of passive transfer of maternal immunoglobulin. On day 10, the calf received a transfusion of concentrated immunoglobulin extracted and concentrated from the cow's previously banked plasma. On day 13, the calf developed a urinary tract infection, as diagnosed by white blood cells and bacteria in the urine. Following immunoglobulin administration and antibiotic therapy, clinical signs slowly resolved and the calf gained weight. The cow passed the fetal membranes during parturition, but the placenta was retained. Despite prophylactic systemic antibiotics and vaginal flushing, the cow became depressed and developed a leukocytosis and an anemia. A mucopurulent vaginal discharge and ventral edema were noted on day 3, and milk production was minimal. Because decreased milk production has been reported as a common sequel to retained placenta, efforts were focused on removing the placenta. Intermittent oxytocin therapy on days 2-14 did not result in expulsion of the placenta and produced only transient abdominal contractions and minimal increases in milk letdown. On day 15, 10 mg estradiol cypionate was administered i.m. followed by 200 IU oxytocin i.v. An additional 10 IU oxytocin was administered i.v. on day 16. The friable placenta was palpable within the vaginal vault on day 17. The remaining placenta was removed by gentle traction applied by a modified weighted blood pressure cuff. Once the placenta was removed, the cow's clinical problems slowly resolved and the calf continued to gain weight.
Old World frugivorous bats have been identified as natural hosts for emerging zoonotic viruses of significant public health concern, including henipaviruses (Nipah and Hendra virus), Ebola virus, and Marburg virus. Epidemiological studies of these viruses in bats often utilize serology to describe viral dynamics, with particular attention paid to juveniles, whose birth increases the overall susceptibility of the population to a viral outbreak once maternal immunity wanes. However, little is understood about bat immunology, including the duration of maternal antibodies in neonates. Understanding duration of maternally derived immunity is critical for characterizing viral dynamics in bat populations, which may help assess the risk of spillover to humans. We conducted two separate studies of pregnant Pteropus bat species and their offspring to measure the half-life and duration of antibodies to 1) canine distemper virus antigen in vaccinated captive Pteropus hypomelanus; and 2) Hendra virus in wild-caught, naturally infected Pteropus alecto. Both of these pteropid bat species are known reservoirs for henipaviruses. We found that in both species, antibodies were transferred from dam to pup. In P. hypomelanus pups, titers against CDV waned over a mean period of 228.6 days (95% CI: 185.4-271.8) and had a mean terminal phase half-life of 96.0 days (CI 95%: 30.7-299.7). In P. alecto pups, antibodies waned over 255.13 days (95% CI: 221.0-289.3) and had a mean terminal phase half-life of 52.24 days (CI 95%: 33.76-80.83). Each species showed a duration of transferred maternal immunity of between 7.5 and 8.5 months, which was longer than has been previously estimated. These data will allow for more accurate interpretation of age-related Henipavirus serological data collected from wild pteropid bats.
Use haematology, biochemistry and protein electrophoresis analyses to establish reference values for, and describe the health status of, wild and captive colonies of critically endangered warru (black-footed rock-wallaby: Petrogale lateralis MacDonnell Ranges race).
Blood samples were taken from warru in three wild colonies (Alalka, Kalka, New Well) in the A nangu Pitjantjatjara Yankunytjatjara Lands in north-west South Australia (SA) and from captive animals at Monarto Zoo, SA. General haematology, serum biochemistry and protein electrophoresis analyses were conducted and results used to establish reference ranges. For the parameters that are indicative of a population's health, comparisons among the study sites were completed using analysis of variance.
General haematology results suggested that warru were not experiencing chronic anaemia and the protein electrophoresis values indicated that colonies were not suffering from population-wide disease. However, the lower superoxide dismutase, retinol, total carotenoids and ascorbic acid values for New Well warru suggested those animals had a lower plane of nutrition than those at Kalka and Alalka. Higher urea concentrations in New Well and Alalka warru could be a reflection of the absence of reliable free water at these sites.
The results have implications for the management of in situ colonies, including potentially using supplementary feeding to improve nutrition, and suggested that these animals were not suffering from disease. The study presents the first blood reference values for P. lateralis and, potentially, a methodology for other threatened species recovery programs to follow to establish the health of their populations.
Watercraft-related trauma is the predominant cause of human-induced mortality in manatees (Trichechus manatus latirostris), a federal- and state-listed endangered species. Pyothorax (documented in this case report) and other secondary infections are common sequelae of inhalation of water and the open wounds caused by boat propellers. These secondary infections can lead to the demise of the animal weeks to months after the traumatic incident when external wounds have healed. Diagnosis of underlying disease on physical examination during capture and restraint can be difficult. Acute phase proteins, including serum amyloid A, fibrinogen, and albumin can be used to diagnose inflammatory disease in manatees and improve quality of medical care and husbandry. We also provide the first report of polycystic kidneys in Sirenians.
C-reactive protein (CRP) is a major, acute-phase protein in dogs; however, there is a need for automated assays to ensure in-time patient monitoring. Three automated immunoturbidimetric assays (Randox, Thermo, and Wako) developed for human beings were evaluated for their ability to detect canine CRP, including method validation, evaluation of diagnostic use, and establishment of exploratory reference intervals. Sera from 36 healthy dogs and 82 diseased dogs were included for method comparison with the enzyme-linked immunosorbent assay (ELISA; Tridelta) serving as the reference method. A nonparametric estimate of the 1-sided 95% reference interval was established (n = 36). Precision study revealed good intra-assay coefficients of variation (CVs) of 1-10%, 0-9%, and 2-13% for the Randox, Thermo, and Wako assays, respectively. Interassay CVs were 18%, 24%, and 19% respectively. Because of a low linear range, the Thermo test was considered unsuitable for use with canine specimens. No significant differences were present between the results obtained with the Randox and Wako assays with CRP concentrations less than 15 mg/l; however, median CRP results differed significantly between the Thermo test and the ELISA (P = 0.03). Bland-Altman analysis detected a proportional bias of 0.28, -0.59, and 0.61 mg/l for the Randox, Thermo, and Wako assays, respectively. For all tests, median CRP values were significantly different between healthy dogs and dogs with neoplasia. The upper limit of the reference intervals were 8.2 and 9.9 mg/l for the Randox and Wako assays, respectively. In contrast to the Thermo test, the Randox and Wako assays were suitable for detection of abnormally high canine CRP concentrations; however, improvement of assay precision and evaluation of accuracy are warranted before their clinical use with canine specimens.
Background:
Maternal antibodies are believed to play an integral role in protecting immunologically immature wild-passerines from environmental antigens. This study comprehensively examines the early development of the adaptive immune system in an altricial-developing wild passerine species, the house sparrow (Passer domestics), by characterizing the half-life of maternal antibodies in nestling plasma, the onset of de novo synthesis of endogenous antibodies by nestlings, and the timing of immunological independence, where nestlings rely entirely on their own antibodies for immunologic protection.
Methodology/principal findings:
In an aviary study we vaccinated females against a novel antigen that these birds would not otherwise encounter in their natural environment, and measured both antigen-specific and total antibody concentration in the plasma of females, yolks, and nestlings. We traced the transfer of maternal antibodies from females to nestlings through the yolk and measured catabolisation of maternal antigen-specific antibodies in nestlings during early development. By utilizing measurements of non-specific and specific antibody levels in nestling plasma we were able to calculate the half-life of maternal antibodies in nestling plasma and the time point at which nestling were capable of synthesizing antibodies themselves. Based on the short half-life of maternal antibodies, the rapid production of endogenous antibodies by nestlings and the relatively low transfer of maternal antibodies to nestlings, our findings suggest that altricial-developing sparrows achieve immunologic independence much earlier than precocial birds.
Conclusions/significance:
To our knowledge, this is the first in depth analyses performed on the adaptive immune system of a wild-passerine species. Our results suggest that maternal antibodies may not confer the immunologic protection or immune priming previously proposed in other passerine studies. Further research needs to be conducted on other altricial passerines to determine if the results of our study are a species-specific phenomenon or if they apply to all altricial-developing birds.
The acute phase response is a complex systemic early-defense system activated by trauma, infection, stress, neoplasia, and inflammation. Although nonspecific, it serves as a core of the innate immune response involving physical and molecular barriers and responses that serve to prevent infection, clear potential pathogens, initiate inflammatory processes, and contribute to resolution and the healing process. Acute phase proteins, an integral part of the acute phase response, have been a focus of many applications in human diagnostic medicine and recently have been identified in common animal species. Potential applications to diagnosis, prognosis, assessment of animal health, and laboratory animal welfare are readily apparent.
Between 1994 and 1996, three elephants from an exotic animal farm in Illinois died of pulmonary disease due to Mycobacterium tuberculosis. In October 1996, a fourth living elephant was culture-positive for M. tuberculosis. Twenty-two handlers at the farm were screened for tuberculosis (TB); eleven had positive reactions to intradermal injection with purified protein derivative. One had smear-negative, culture-positive active TB. DNA fingerprint comparison by IS6110 and TBN12 typing showed that the isolates from the four elephants and the handler with active TB were the same strain. This investigation indicates transmission of M. tuberculosis between humans and elephants.
The deaths of two Asian elephants (Elephas maximus) in August 1996 led the United States Department of Agriculture to require the testing and treatment of elephants for tuberculosis. From August 1996 to September 1999. Mycobacterium tuberculosis infection was confirmed by culture in 12 of 118 elephants in six herds. Eight diagnoses were made antemortem on the basis of isolation of M. tuberculosis by culture of trunk wash samples; the remainder (including the initial two) were diagnosed postmortem. We present the case histories, epidemiologic characteristics, diagnostic test results, and therapeutic plans from these six herds. The intradermal tuberculin test, enzyme-linked immunosorbent assay serology, the blood tuberculosis test, and nucleic acid amplification and culture are compared as methods to diagnose M. tuberculosis infection in elephants.
The body's early defence in response to trauma, inflammation or infection, the acute phase response, is a complex set of systemic reactions seen shortly after exposure to a triggering event. One of the many components is an acute phase protein response in which increased hepatic synthesis leads to increased serum concentration of positive acute phase proteins. The serum concentration of these acute phase proteins returns to base levels when the triggering factor is no longer present. This paper provides a review of the acute phase proteins haptoglobin, C-reactive protein and serum amyloid A and their possible use as non-specific indicators of health in large animal veterinary medicine such as in the health status surveillance of pigs at the herd level, for the detection of mastitis in dairy cattle and for the prognosis of respiratory diseases in horses.
The African elephant reproduces so efficiently in the wild that overpopulation is now a serious problem in some game parks in Zimbabwe, Botswana and South Africa. The female reaches puberty between 10 and 12 years of age in the wild and, when in captivity, shows oestrous cycles of 14-15 weeks duration. She readily conceives a singleton in the wild yet her uterus has the capacity for twins. She shows a gestation length of 22 months and, in the wild, shows a population density and feed dependent intercalving interval of 4-8 years. The trophoblast erodes the lumenal epithelium of the endometrium and stimulates upgrowths of blood vessel-containing stromal villi, which develop eventually into the broad, tightly folded lamellae of the zonary, endotheliochorial placenta. Significant quantities of leaked maternal erythrocytes and ferric iron are phagocytosed by specialized trophoblast cells in the haemophagous zones at the lateral edges of the placental band. Although the placenta itself is endocrinologically inert, the foetal gonads, which enlarge greatly during the second half of pregnancy can synthesize 5alpha-dihydryoprogesterone and other 5alpha pregnane derivatives from cholesterol and pregnenolone. These products may synergize with progestagens secreted by the 2-8 large corpora lutea which are always present in the maternal ovaries throughout gestation to maintain the pregnancy state.
Grant's zebra (Equus burchelli) are commonly kept in zoos and are subject to routine health monitoring and research studies. Recently, assays for acute phase proteins (APP) have been described in many wildlife species, and specific assays for serum amyloid A (SAA) have been well validated and studied in horses (Equus ferus caballus), in which it serves as a major APP. In the present study, serum samples from 26 Grant's zebra were subject to analysis by using assays for SAA, haptoglobin (HP), and protein electrophoresis. Reference intervals were calculated by using the robust method: SAA 1.8-31.4 mg/L and HP 0.37-1.58 mg/ml. Significant differences in SAA and HP were observed in clinically abnormal zebra; in some cases, these differences were marked and were noted in the absence of abnormal values for protein electrophoretic fractions. These data indicate that APP may be a valuable and sensitive tool in monitoring inflammation in this species.
The acute phase protein serum amyloid A (SAA) has been previously shown to have value as a biomarker of inflammation and infection in many species, including manatees (Trichechus manatus latirostris). In the current study, results from an automated assay for SAA were used in a rehabilitation setting. Reference intervals were established from clinically normal manatees using the robust method: 0-46 mg/L. More than 30-fold higher mean SAA levels were observed in manatees suffering from cold stress and boat-related trauma. Poor correlations were observed between SAA and total white blood count, percentage of neutrophils, albumin, and albumin/globulin ratio. A moderate correlation was observed between SAA and the presence of nucleated red blood cells. The sensitivity of SAA testing was 93% and the specificity was 98%, representing the highest combined values of all the analytes. The results indicate that the automated method for SAA quantitation can provide important clinical data for manatees in a rehabilitation setting.
Infection of Asian elephants (Elephas maximus) with elephant endotheliotropic herpesvirus (EEHV) can be associated with rapid, lethal hemorrhagic disease and has been documented in elephant herds in human care and in the wild. Recent reports describe real-time quantitative polymerase chain reaction (qPCR) assays used to monitor clinically ill elephants and also to detect subclinical EEHV1 infection in apparently healthy Asian elephants. Acute phase proteins have been demonstrated to increase with a variety of infectious etiologies in domesticated mammals but have not yet been described in elephants. In addition, the immune response of Asian elephants to EEHV1 infection has not been described. In this study, whole blood and trunk wash samples representing repeated measures from eight elephants were examined for the presence of EEHV1 using a qPCR assay. Elephants were classified into groups, as follows: whole blood negative and positive and trunk wash negative and positive. Serum amyloid A (SAA) and haptoglobin (HP) levels were compared between these groups. A significant difference in SAA was observed with nearly a threefold higher mean value during periods of viremia (P=0.011). Higher values of SAA were associated with >10,000 virus genome copies/ml EEHV1 in whole blood. There were no significant differences in HP levels, although some individual animals did exhibit increased levels with infection. These data indicate that an inflammatory process is stimulated during EEHV1 viremia. Acute phase protein quantitation may aid in monitoring the health status of Asian elephants.
Objective:
To compare serum amyloid A (SAA) concentration, plasma fibrinogen concentration, total WBC count, and serum albumin-to-globulin concentration ratio (A:G ratio) in clinically normal (CN) and clinically abnormal (CA) horses.
Design:
Prospective cohort study.
Animals:
111 CN horses and 101 CA horses hospitalized at a specialty clinical practice.
Procedures:
Shortly after admission, a blood sample (20 mL) was collected from each horse for a CBC, serum protein electrophoresis, and determination of plasma fibrinogen concentration; SAA concentration was assessed with a previously validated immunoturbidometric assay. Similar testing of a subset of CA horses was conducted at various points during treatment.
Results:
Total WBC count, A:G ratio, and SAA concentration were determined for all 212 horses; data regarding plasma fibrinogen concentration were available for 127 horses (of which 47 were CN and 80 were CA). Median SAA concentration, total WBC count, and plasma fibrinogen concentration and mean A:G ratio differed significantly between CN horses and CA horses. Correlations between these variables were poor to weak. For discrimination of CN horses from CA horses, the SAA assay had sensitivity of 53% and specificity of 94% (diagnostic accuracy, 75%); for the other assessments, accuracy ranged from 59% to 62%. Repeated assessment of SAA concentration in some CA horses revealed a gradual return to normal concentrations.
Conclusions and clinical relevance:
Results indicated that assessment of SAA concentration can provide valuable information regarding the clinical state of horses and may be more useful for patient monitoring and as a prognostic indicator than are traditional markers of inflammation.
SUMMARY Mycobacterium tuberculosis is primarily a pathogen of humans. Infections have been reported in animal species and it is emerging as a significant disease of elephants in the care of humans. With the close association between humans and animals, transmission can occur. In November 2010, a clinically healthy Asian elephant in an Australian zoo was found to be shedding M. tuberculosis; in September 2011, a sick chimpanzee at the same zoo was diagnosed with tuberculosis caused by an indistinguishable strain of M. tuberculosis. Investigations included staff and animal screening. Four staff had tuberculin skin test conversions associated with spending at least 10 hours within the elephant enclosure; none had disease. Six chimpanzees had suspected infection. A pathway of transmission between the animals could not be confirmed. Tuberculosis in an elephant can be transmissible to people in close contact and to other animals more remotely. The mechanism for transmission from elephants requires further investigation.
Background:
Captive elephants infected with tuberculosis are implicated as an occupational source of zoonotic tuberculosis. However, accurate estimates of prevalence and incidence of elephant tuberculosis from well-defined captive populations are lacking in the literature. Studies published in recent years contain a wide range of prevalence estimates calculated from summary data. Incidence estimates of elephant tuberculosis in captive elephants are not available.
Objective:
This study estimated the annual point prevalence, annual incidence, cumulative incidence, and incidence density of tuberculosis in captive elephants within the USA during the past 52 years.
Animals and methods:
We combined existing elephant census records from captive elephants in the USA with tuberculosis culture results obtained from trunk washes or at necropsy. This data set included 15 years where each elephant was screened annually.
Results:
Between 1960 and 1996, the annual point prevalence of tuberculosis complex mycobacteria for both species was 0. From 1997 through 2011, the median point prevalence within the Asian elephant population was 5.1%, with a range from 0.3% to 6.7%. The incidence density was 9.7 cases/1000 elephant years (95% CI: 7.0-13.4). In contrast, the annual point prevalence during the same time period within the African elephant population remained 0 and the incidence density was 1.5 cases/1000 elephant years (95% CI: 0.7-4.0).
Conclusions:
The apparent increase in new cases noted after 1996 resulted from a combination of both index cases and the initiation of mandatory annual tuberculosis screening in 1997 for all the elephants. This study found lower annual point prevalence estimates than previously reported in the literature. These discrepancies in prevalence estimates are primarily due to differences in terminology and calculation methods. Using the same intensive testing regime, the incidence of tuberculosis differed significantly between Asian and African elephants.
Clinical importance:
Accurate and species specific knowledge of prevalence and incidence will inform our efforts to mitigate occupational risks associated with captive elephants in the USA.
Acute phase proteins (APPs) have been studied in many companion and large animals and have been reported to have a differential sensitivity to traditional markers of inflammation. Studies have been performed indicating the conservation of these proteins as well as the application and cross-reactivity of automated assays among different species, but few reports have detailed APPs in marine mammal species. In the present study, automated assays were utilized to generate reference intervals for C-reactive protein, haptoglobin, and serum amyloid A using 44 serum samples from healthy Atlantic bottlenose dolphins (Tursiops truncatus). A total of 25 samples were obtained from dolphins under human care and 19 samples were obtained from free-ranging dolphins. Mild yet statistically significant differences were observed in levels of haptoglobin and serum amyloid A between these groups. The reference intervals from the combined groups were as follows: C-reactive protein 3.1-19.7 mg/l, haptoglobin 0-0.37mg/ml, and serum amyloid A 17.5-42.9 mg/l. These baseline data should provide an important foundation for future studies of the application of APP quantitation in monitoring the health and stressors of dolphins under human care and with live capture of free-ranging dolphins.
1.1.Elephant blood clots rapidly and the clots retract only slightly. The partial thromboplastin time is faster than human and most coagulation factors are much higher when assayed in systems designed for human plasma.2.2.Thrombin time was long but antithrombin III was essentially absent.3.3.Platelet counts were in the high-normal human range and aggregation with ADP and collagen was less marked than in human platelet rich plasma.4.4.Red cells were large, discoid in shape, but fewer in number than in human blood.5.5.Leukocytes were present in greater numbers than in human and showed some morphological differences.6.6.Some biochemical and protein differences were also observed.
1.1. Electrophoresis of blood serum of the African elephant separated 5 protein components designated albumin, α-, β1-, β2 and γ-glubulin. Quantitation of these protein fractions showed that there were significant seasonal and geographical variations in the levels.2.2. Similar seasonal or geographical variations were found in the levels of urea, thyroxine, T3 uptake and creatinine.3.3. The significance of these changes is discussed.
In a clinical setting, especially with species of special interest, it is important to use all clinical pathology testing options for general health monitoring and diagnosis. Protein electrophoresis (EPH) has previously been shown to be an important adjunct tool in veterinary medicine. Serum samples from 18 free-ranging and 12 zoo-based koalas (Phascolarctos cinereus) were subject to EPH analysis. Significant differences were found between the two groups for the following values: total protein, albumin, beta globulins, and albumin-globulin ratio (P < 0.05). By using the combined data, the minimum-maximum values for the EPH fractions were as follows: total protein 5.0-7.8 g/dl, albumin 2.8-4.7 g/dl, alpha-1 globulins 0.5-1.1 g/dl, alpha-2 globulins 0.3-0.7 g/dl, beta globulins 0.4-1.0 g/dl, gamma globulins 0.2-1.0 g/dl, and albumin-globulin ratio 1.0-2.1.
Acute phase proteins (APP) were first identified in the early 1900s as early reactants to infectious disease. They are now understood to be an integral part of the acute phase response (APR) which is the cornerstone of innate immunity. APP have been shown to be valuable biomarkers as increases can occur with inflammation, infection, neoplasia, stress, and trauma. All animals--from fish to mammals--have demonstrable APP, but the type of major APP differs by species. While the primary application of these proteins in a clinical setting is prognostication, studies in animals have demonstrated relevance to diagnosis and detection and monitoring for subclinical disease. APP have been well documented in laboratory, companion, and large animals. With the advent of standardized and automated assays, these biomarkers are available for use in all fields of veterinary medicine as well as basic and clinical research.
The use of major acute-phase proteins (APPs) for assessment of health and disease in companion animals has increased within the last decade because of increased knowledge in the field and increased access to appropriate assay systems for detection of relevant APPs, which are highly species specific. Despite evidence being restricted almost solely to proven excellent overlap performance of these markers in detecting inflammatory activity, clinically relevant studies at higher evidence levels do exist. The available body of literature shows a clear, but seemingly untapped, potential for more extended routine clinical use of major APP testing in companion animal medicine.
Acute phase proteins (APPs) have been used as biomarkers of inflammation, infection and trauma for decades in human medicine but have been relatively under-utilised in the context of veterinary medicine. However, significant progress has been made in the detection, measurement and application of APPs as biomarkers in both companion and farm animal medicine over recent years. In the dog, C-reactive protein, haptoglobin and serum amyloid A have been identified as significant diagnostic 'markers' of steroid-responsive meningitis-arteritis, while in cats and cattle haptoglobin and alpha(1) acid glycoprotein and haptoglobin and serum amyloid A have proved valuable biomarkers of disease, respectively. In dairy cattle, haptoglobin and a mammary-associated serum amyloid A3 isoform, produced by the inflamed mammary gland during episodes of mastitis, have great potential as biomarkers of this economically important disease. Understanding the use of APP as biomarkers of inflammatory conditions of domestic animals has expanded significantly over recent years, and, with the insights provided by ongoing research, it is likely that these compounds will be increasingly used in the future in the diagnosis and prognosis of both companion and farm animal disease.
The serum concentration of acute phase proteins (APPs) increases dramatically in response to inflammation and tissue injury. APPs are clinically useful in a range of domesticated mammals; however, knowledge is limited in nondomesticated mammals. The detective ability of two assays for each of three potential APPs--serum amyloid A (SAA), C-reactive protein (CRP), and haptoglobin (Hp)--was evaluated in eight species. For SAA, a turbidimetric immunoassay (TIA) demonstrated significant detective abilities in the Asian elephant (Elaphas maximus), impala (Aepyceros melampus), musk ox (Ovibos moschatus), and chimpanzee (Pan troglodytes), as did an SAA enzyme-linked immunosorbent assay (ELISA) in the impala. For CRP, both TIA and ELISA had significant detective abilities in the chimpanzee. For Hp, a colorimetric assay demonstrated significant detective abilities in impala, musk ox, sitatunga (Tragelaphus spekeii), and chimpanzee, as did the Hp ELISA in the impala, musk ox, and sitatunga. In conclusion, these results suggest that assays for detection of relevant APPs in several nondomesticated animals are available.
Inflammation induces dramatic changes in the biosynthetic profile of the liver, leading to increased serum concentrations of positive acute phase (AP) proteins and decreased concentrations of negative AP proteins. Serum amyloid A (SAA) and the pentraxins C-reactive protein (CRP) and serum amyloid P component (SAP) are major AP proteins: their serum levels can rise by 1000-fold, indicating that they play a critical role in defense and/or the restoration of homeostasis. We have cloned SAA and a SAP-like pentraxin from salmonid fish species. The salmonid SAA shares approximately 70% amino acid identity with mammalian AP SAA. When salmonids are challenged with an AP stimulus, i.e., Aeromonas salmonicida, SAA responds dramatically as a major AP reactant. The salmonid pentraxin shows approximately 40% amino acid identity to both mammalian SAP and CRP. Evolutionary analysis suggests the presence of only a single such protein in teleosts and lower animal species. Surprisingly, the salmonid pentraxin behaves as a negative AP reactant, reminiscent of the SAP-like Syrian hamster female protein, in that hepatic mRNA concentrations decline to 50% of prestimulus levels. This study reinforces the hypothesis that SAA induction is an essential and universal feature of the vertebrate AP response and that it represents part of an ancient host defense system. Conversely, the species-dependent heterogeneity of pentraxin expression during the vertebrate AP response supports the possibility that its most important ancestral (and perhaps present) function is not related to its AP behavior.
C-reactive protein (C-RP) was purified from harbor seal (Phoca vitulina) serum by calcium dependant phosphoryl-choline and protein A affinity chromatography. Polyacrylamide gel electrophoresis under reducing conditions revealed a single protein moiety with a molecular weight of approximately 25 kDa. An internal peptide derived from this purified protein was subjected to N-terminal amino acid sequencing. A high amino acid sequence similarity was obtained with other published mammalian C-RP molecules confirming that the purified protein was a C-RP homologue. Eight specific monoclonal antibodies (P13, P51, P87, P101, P106, P130, P157 and P219) were raised against this purified protein. All 8 monoclonal antibodies immunoblotted with the 25 kDa C-RP subunit under reducing conditions. A competitive immunoassay was developed identifying elevated C-RP concentrations in harbor seal serum samples with clinical evidence of inflammatory disease. Application of this immunoassay for the measurement C-RP may provide valuable information for the clinical assessment of harbor seal health.
Serum C-reactive protein (CRP) is an acute phase marker in dogs that is useful for the diagnosis and monitoring of inflammatory disease. Rapid, reliable, and automated assays are preferable for routine evaluation of canine serum CRP concentration.
The aim of this study was to evaluate whether canine serum CRP concentration could be measured reliably using an automated turbidometric immunoassay (TIA) designed for use with human serum.
A commercially available TIA for human serum CRP (Bayer, Newbury, UK) was used to measure canine serum CRP concentration. Cross-reactivity of antigen was evaluated by the Ouchterlony procedure. Intra- and interassay imprecision was investigated by multiple measurements on canine serum samples and serum pools, respectively. Assay inaccuracy was investigated by linearity under dilution and comparison of methodologies (canine CRP ELISA, Tridelta Development Ltd, Kildare, UK). Then the assay was applied to serum samples from 14 clinically healthy dogs, 11 dogs with neoplasia, 13 with infections, 8 with endocrine or metabolic diseases, and 10 with miscellaneous diseases.
Cross-reactivity between canine serum CRP and the anti-human CRP antibody was found. Intra- and interassay imprecision ranged from 5.2% to 10.8% and 3.0% to 10.2%, respectively. Serum CRP concentration was measured in a linear and proportional manner. There was no significant disagreement and there was linear correlation of the results in the comparison of methodologies, except for a slight proportional discrepancy at low CRP concentrations (<10 microg/mL). Dogs with infections had a significantly higher concentration of serum CRP than did all other dogs, and dogs with neoplasia had a significantly higher concentration of serum CRP than did clinically healthy dogs.
Canine serum CRP concentration can be measured reliably using the commercially available TIA designed for human CRP.
The acute phase proteins (APP) are a group of blood proteins that contribute to restoring homeostasis and limiting microbial growth in an antibody-independent manner in animals subjected to infection, inflammation, surgical trauma or stress. In the last two decades, many advances have been made in monitoring APP in both farm and companion animals for clinical and experimental purposes. Also, the mechanism of the APP response is receiving attention in veterinary science in connection with the innate immune systems of animals. This review describes the results of recent research on animal APP, with special reference to their induction and regulatory mechanisms, their biological functions, and their current and future applications to veterinary diagnosis and animal production.
Afrotheria, one of four mammalian superorders, comprises elephants, sea cows, hyraxes, aardvark, elephant shrews, tenrecs and golden moles. Their placentas either form an equatorial band or are discoid in shape. The interhemal region, separating fetal and maternal blood, is endotheliochorial in elephants, aardvark and possibly the sea cows, but hemochorial in the remaining orders. There is a secondary epitheliochorial placenta in elephant shrews while a similar structure in tenrecs erodes maternal tissues. Specialized hemophagous regions are a striking characteristic of some of these placentas yet absent in hyraxes, elephant shrews, and golden moles. It is possible that the common ancestor of the Afrotheria had an endotheliochorial placenta. Establishment of a hemochorial condition, as seen in rock hyraxes, elephant shrews, tenrecs, and golden moles, would be a more recent development. The elephant, manatee, and aardvark all have circumferential placentas. Thus the formation of a discoid placenta with a more or less extensive secondary placenta in elephant shrews and tenrecs would also be a derived state.
The acute phase response is a nonspecific inflammatory reaction of the host that occurs shortly after any tissue injury. The response includes changes in the concentration of plasma proteins called acute phase proteins (APPs), some of which decrease in concentration (negative APPs), such as albumin or transferrin, and others of which increase in concentration (positive APPs), such as C-reactive protein, serum amyloid A, haptoglobin, alpha-1-acid glycoprotein, and ceruloplasmin. Most positive APPs are glycoproteins synthesized mainly by hepatocytes upon stimulation by proinflammatory cytokines and released into the bloodstream. The acute phase response and clinical application of monitoring APPs in dogs and cats are reviewed in this article, including biochemical characteristics, assays developed for each individual APP, and preanalytic and analytic factors influencing APP results that should be taken into account for proper and adequate clinical interpretation. In addition, the diagnostic use of APPs and their possible application in monitoring treatment, which can be considered one of the most interesting and promising practical applications of these proteins, will be discussed. Finally, challenges and future developments of APPs in dogs and cats will be considered, because it is expected that new and cheaper automated assays for determination of the main APPs in small animals will contribute to a wider use of these proteins as biomarkers of infection and inflammatory lesions.
The aim of the present study was to evaluate whether equine serum amyloid A (SAA) concentrations could be measured reliably with a turbidometric immunoassay (TIA) developed for use with human serum. Intra- and inter-assay imprecision were evaluated by multiple measurements on equine serum pools. Assay inaccuracy was determined by linearity under dilution. The assay was subsequently used for measuring SAA concentrations in clinically healthy horses, horses with inflammatory diseases, horses with non-inflammatory diseases, and in horses before and after castration. In pools with low, intermediate and high SAA concentrations, the intra-assay imprecisions were 24.4%, 1.6% and 2.1%, and the inter-assay imprecisions were 33.2%, 4.6% and 6.5%. Slight signs of inaccuracy were observed, but these inaccuracies were negligible when considering the large dynamic range of the SAA response. The assay was able to detect the expected difference in SAA levels in different groups of horses. It was also able to demonstrate the expected dynamic changes in SAA after castration. In conclusion, equine SAA concentrations can be measured reliably using the TIA designed for human SAA.
Abnormal physiological conditions and diseases can change the concentrations of enzymes, metabolites, and minerals in the body. Serum chemistry information may thus be indicative of a specific disease; interpretation of such information requires knowledge of serum chemistry reference intervals from a seemingly healthy population of the species.
The aim of this study was to obtain serum chemistry reference intervals for a population of white whales.
Blood samples were collected from 21 free-ranging white whales (beluga; Delphinapterus leucas). The whales were live-captured in nets during 1996-2001 in Storfjorden, Van Mijenfjorden, and Van Keulenfjorden (Svalbard, Norway). While the whales were briefly physically restrained, blood was collected from the caudal vein into vacuum tubes without anticoagulant. The blood was left to clot for 4-6 hours before serum was obtained by centrifugation. The serum samples were then kept at -20 degrees C until analysis. Enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase [ALP], creatine kinase, lactate dehydrogenase [LDH], amylase, lipase), metabolites (urea, creatinine, bilirubin, cholesterol, triglycerides, nonesterified fatty acids, glucose), and minerals (calcium, phosphate, magnesium, sodium, potassium, chloride) were analyzed in an Advia 1650 System (Bayer, Tarrytown, NY, USA). Cortisol was analyzed in an Immulite One system (Diagnostic Products Corporation, Los Angeles, CA, USA). The major blood proteins (albumin and globulins) were separated by gel electrophoresis in a Beckman Paragon electrophoresis system (Beckman Coulter, Inc., Fullerton, CA, USA).
Serum values for all analytes were reported as median and range, and reference intervals were calculated as 10-90th percentiles. Activities of ALP and LDH and cortisol concentration were higher, and protein and bilirubin concentrations were lower compared with those previously reported for white whales from Canada; remaining results were strikingly similar in these 2 white whale populations.
These data provide valuable serum chemistry reference intervals for future health assessments of white whales in Svalbard and other white whale populations, as well as captive individuals.
Proteins, proteomics, and the dysproteinemias In: Clinical biochemistry of domestic animals
- Pd Eckersall
- Jj Kaneko
- Jw Harvey
- Ml Bruss
Eckersall PD: 2008, Proteins, proteomics, and the dysproteinemias. In: Clinical biochemistry of domestic animals,
ed. Kaneko JJ, Harvey JW, Bruss ML, 6th ed., pp. 117–155.
Academic Press, San Diego, CA.