The results of the investigations carried out allow us to establish that the deaths that occurred for years in that paddock and the investigations by us are due to cyanide poisoning, the ingestion of fruits and seeds of Pouteria campechiana (Canistel) by the heifers and that it was corroborated the presence of Chronogenic Glucosides in this plant and that as Runnells and Monlux (1975) suggest, these are hydrolyzed in the root by the enzymes of the microflora.
1. Introduction
Accidental poisoning is of greater importance in animals if one considers the multitude of opportunities they have in ingesting toxic substances. Among them, Ruminants, due to their characteristics of consuming large volumes of food, together with the complex mechanical and biochemical processes that occur in their stomachs during digestion, have a higher potential risk.
Poisons are one of the most important causes of mortality in our country and in any livestock farming system, causing great economic losses due to the high morbidity and mortality that they cause, as well as the expenses on medications that, depending on their recovery, it incurs and costs, generally high, of the diagnostic techniques for the detection and confirmation of the acting substance.
In a study of poisonings diagnosed in the country by the Central Laboratory of Veterinary Diagnostics for 17 years, the natural toxins contained in toxic plants and grasses originated 66.6% of the poisonings and 5.0% were Cyanogenic Glycosides, (Mirtha Rivero, 1993).
Hydrocyanic Acid or Prussic Acid (HCN) is one of the most toxic and rapidly active poisons.
The most important source of Hydrocyanic Acid for animals is by far of plant origin. Many plant species contain free or more frequently Hydrocyanic Acid, in the form of Cyanogenic Glycosides, organic compounds that contain a sugar and are capable of releasing cyanide by hydrolysis.
Glucoside is not toxic by itself, but if it comes in contact with the appropriate enzyme it breaks down and releases Hydrogen Cyanide.
Glycosides are not toxic until Hydrocyanic Acid is released from them by the activity of microorganisms in the belly, or by the action of an enzyme contained in this or other plants. For this reason, photogenic cyanide poisoning occurs mainly in ruminants (Jubb and Kennedy, 1974).
Not always the depth of the Clinical-Epizootiological surveys and the anamnesis is capable of guiding the Toxicological diagnosis, so that as in the cases that we will expose, for years and in seasonal periods high levels of deaths occurred in which the Business Veterinary Service and the Diagnostic Group, which included the Laboratories, could not reach conclusions; breeders taking the measure of not allowing access to these paddocks of cows, not ruling out the suspicion that there was some intention in these cases.
Our work aims to report for the first time the poisoning from the ingestion of Pouteria campechiana (Canistel) in ruminants that were grazing; describe the steps of the investigation, as well as the botanical and physiological conditions that caused these deaths. Describe the symptoms and injuries observed and recommend the use of a simple and low-cost Clinical Laboratory technique to guide this diagnosis.
At the same time alert the centers that work in the propagation and rescue of this fruit tree about the risk that it represents for ruminants.
Botany
The Canistel, whose scientific name is Pouteria campechiana, Lucuma nervosa or Lucuma campechiana, belongs to the Ebenales order and to the Sapotaceae family, it is a tree sometimes up to 25 m., Commonly smaller; elliptical and lanceolate leaves, (Amaro, 2002; Herrera, 2006; and Dec. 2008).
The tree is neither tall nor common; It is medium in size and made of good yellowish pink wood. It produces the fruit of the same name, something similar to the Sapote snake, subglobose or piriform fruit of up to 7 cm., green to brownish, yellow orange in maturity and inside as an egg yolk, which according to Carmen Cabrera (2003), Canistel has a high content of vitamin A, which is appreciated for its intense yellow color. Its dry matter reaches 40% and its mass is compact. Its seed is small and the external color is similar to that of Mamey, Canistel's ice cream is delicious, (Rafaela Manso, 2003).
It has been described as edible and similar to mango since the middle of the last century and as part of the flora of Cuba it was pointed out that it was sometimes cultivated in the former provinces of Las Villas, Habana and Isla de Pinos, (Sauget and Lioguer, 1957; Roig, 1962). It is also distributed in Florida and from Mexico to Panama.
Herrera (2006), in his Artificial Classification System of synatrophic Magnoliats of Cuba, classifies it as a species of allochthonous origin (Exotic, Foreign) introduced with or without the intention of man, who generally can almost always live in open spaces due to atrophic action.
When evaluating Fruit trees in 38 Conucos from 3 regions of Cuba, Canistel was only found in the west of the country and the rest of the species were more abundant in the central region (Fuentes, 2005). Also the work of the Urban Agriculture in the municipality of Cruces only found two Canistel plants in a plot and one of them in production (Hernández and Socorro, 2004).
The Pouteria campechiana, its leaves are reported as santeria herbs (OBATALA) to contain bleeding and blood sputum.
This fruit is being developed by the FAO Regional Office in Organic Agriculture, and Canistel is cited as an example of native and local varieties in order to recover them and exploit their potential (Leonor Castineira and Shagarodosky, 2002).
Due to the total absence in the market of Canistel together with the Níspero, Chirimoya, Anón and guanábana they are part of what could be called exotic fruits on the national scene, but efforts of agriculture together with the Non-Governmental Society Antonio Núñez Jiménez have tried to reverse this situation, (Rafaela Manso, 2004).
2. Materials and Methods:
The Epizootiological study and the review of all cases of sudden death occurred in the "Las Auras" Development Center, belonging to the Yayabo Livestock Company, from the municipality of Sancti-Spiritus, were carried out from 1985 to 1993 and in which no conclusions had been reached, necropsies were performed at the site of dead and slaughtered animals to undergo microscopic and microscopic investigations of the samples with lesions, which were previously fixed in 10% Neutral Formol, subjected to the Freezing and Paraffin techniques. and stained with Hematoxylin-Eosin.
Bacteriological, Bromatological investigations of food and mineral salts and common salt were carried out; Toxicological organs ruminal content and muscles for the detection of nitrates and nitrites, toxic alkaloids.
The Picrate Test was mounted, applying it to the ruminal content and the seeds detected in it, as well as a Biological Test was carried out on 12 white Bal / C mice of which 6 were injected intraraperitoneally with 90 mg. POUTERIA Campechana (Canistel) seeds maceration, in doses from 0.3ml to 2.5ml, while the controls were administered distilled water by a similar route, and the two groups maintained similar conditions in the Bioterio of the Provincial Diagnostic Laboratory.
Toxicology samples that included blood were also sent to the Toxicology Department of the Villa Clara Provincial Laboratory in order to determine Phosphorous Organs.
A botanical survey was carried out in the affected paddocks with the support of the Experimental Station of Pastures and Forages of S, Spiritus, which included an in situ tracking of the local plants, classifying them as toxic or non-toxic.
Case report:
The "Las Auras" Development Plan, belonging to the Yayabo Livestock Company, in the Sancti-Spíritus municipality, is located in a pre-mountainous area, difficult to access, but endowed with good natural grass that allowed it to be used for development of Holstein x Zebu crossbred heifers that are extracted towards the dairy farms already gestated from that unit. Its areas limit in some paddocks with land that is destined for the sowing of various crops (garlic, cucumbers, etc…) and that receive periodic fertilization and fumigation, depending on the type of crops.
For years there have been episodes of "Sudden Deaths", which, given the distance and difficulties of access, had only been able to reach the diagnosis of Intoxication due to the clinical symptoms and injuries observed in the place, but the Laboratory studies due to the lateness of the samples In reaching the same, the signs of autolysis and the negative results of the complementary investigations were not effective, so the use of these pastures by the breeders was avoided due to the high levels of economic losses, underusing areas of good food.
The damages that the heifers when escaping sometimes from the paddocks due to difficulties in the fencing, caused the sowings of the residents of the place allowed not to rule out some intentional fact, so the competent agencies also joined these surveys.
On March 24, 1985 the first sick animals were observed with nervous symptoms appreciated by the main doctor, absence of fever. Treatment is applied to the group of 10 sick heifers, but at 48 hours 10 heifers die, the autopsy was performed by the Provincial Diagnostic Group, observing discrete petechial hemorrhages in the heart and urinary bladder and ecchymosis in livers and muscles.
Samples are taken from the viscera, ruminal content and the foods they consumed, including mineral salts and common salt, undergoing Anatomo-Pathological, Bacteriological, Bromatological and Toxicological investigations with negative results.
For several years, as we pointed out, the use of these paddocks is avoided by ranchers, a common practice among our breeders when they suspect “sudden death”; but the need for good pasture and watering forces them to retake the area and on 03/23/93 it is reported that in the unit there are 15 sick heifers with similar symptoms, so the visit is carried out and the heifers in good nutritional condition were observed and of development, in lateral decubitus, muscular tremors of the head, eyelids, lips, neck, mydriasis and blindness. Meteorism, rectal tenesmus, temperature of 36.5 - 37 ° C and congestive mucosa. Breathing was markedly breathless, bradycardia and blood difficult to coagulate, with a dark tint and in the dark showing moderate brightness.
The pathological lesions present in sacrificed animals were liver degeneration, hepatomegaly and very friable, ecchymosis in the spleen and kidney. The rumen whose content at the beginning had a PH of 11, falls within a few minutes of being in contact with air of 8.5 - 9; Abundant quantities of Pouteria campechiana (Canistel) seeds are seen in different degrees of decomposition and a characteristic bitter almond odor is emitted from the interior.
The brain presented moderate to severe edema and vascular congestion. Microscopic studies of this organ showed perivascular edema, microbleeds, and areas of tenderness in the cerebellum, especially near the Purkinge cell layer. In the cortex, the areas of Malaysia adopt a linear arrangement, there is edema and neuronal degeneration, but without signs of an inflammatory reaction.
Samples of Canistel seeds macerated in the rumen and fresh seeds of the macerated plant and almonds were selected. At 6 hours of incubation at 37 ° C, bright red coloration was observed in the tubes of both fresh and rumen macerated seeds and at 24 hours all the incubated samples were bright red to gray red. At 72 hours all are positive for the presence of cyanide.
The biological test carried out in the white mice caused in the group that received the seed, muscle tremors, dyspnea, hair standing on end, leading to inactivity and paralysis. Those who received 2.5 ml doses presented cyanosis, seizures and death within 2 hours. The rest died in 2-3 hours.
The lesions in the laboratory mice were similar to the older animals, but more congestive. The control group had no symptoms or injuries.
The rest of the investigations carried out were negative for Nitrates, Nitrites, Toxic Alkaloids, Phosphorous Organs, nor are valuable germs isolated.
In the botanical survey carried out in the area in question, no edible toxic plants were detected by the heifers and capable of producing similar pictures, only one specimen of the Canistel tree was observed, in an area where there was obviously a home and where at the time of Search found large amounts of the ripe fruit in the tree and in the ground with the presence of cows that ingested the same.
Discussion and Conclusions:
The results of the investigations carried out allow us to establish that the deaths that occurred for years in that paddock and the investigations by us are due to cyanide poisoning, the ingestion of fruits and seeds of Pouteria campechiana (Canistel) by the heifers and that it was corroborated the presence of Chronogenic Glucosides in this plant and that as Runnells and Monlux (1975) suggest, these are hydrolyzed in the root by the enzymes of the microflora.
Although the minimum lethal dose of Ac. Free Hydrocyanic and Potassium Cyanide orally is 2.0 - 2.3 mg. From HCN per Kg of live weight in almost all species, it is impossible to indicate with any certainty the toxic dose of cyanide in the form of glucoside, since it varies according to the conditions that occur in plants and it is stated that ruminant poisoning depends of the quantity of ingested plant, of the animal's diet, of the PH of the gastric content and of the concentrations of this free or combined toxic in the ingested plant.
It is generally fast-eating animals that die, and an intake of plant material equivalent to 4 mg / Kg of body weight can be considered fatal. The rapid release of cyanide, with only a short delay in the onset of symptoms, follows the ingestion of Clover or flaxseeds, these statements by Garner and Papworth (1975) coincide with what we observed in natural cases of intoxication. by Canistel and in the Biological Tests carried out since our mice, even in the lowest doses, died within a few hours of receiving the inoculum.
The observation of the brilliance of the red smell of the blood of the intoxicated ruminants is coincident with the differential signs of this poisoning before other types of intoxications with similar symptoms raised by Gibbons and Col, (1972).
It is evident that the lesions observed by us in the nervous system of affected animals is due to the pathological effects of anoxia on nervous tissue with a clear morphological expression of ischemic neuronal necrosis, classified by Jubb and Kennedy, (1974) as histotoxic anoxia, since the venom inhibits the action of the intracellular respiratory enzymes.
Notwithstanding the extensive bibliography consulted in the Botanical and Toxicological order and that some specialists relate more than 59 chronogenic plants, the Poutería Campechirna is not described in the universal literature as a producer of these toxic pictures and if there is a tendency to promote it, therefore that we believe it is important to alert about these negative characteristics in ruminants not only in Cuba but also in the geographical areas of America in which this fruit is distributed.
We must also not forget the importance of a multisectoral approach in this diagnosis and the simple application of the Pierato de Sodio Test, González, (2003), as well as the incorporation of the Biological Test in white mice.
Our results also reduced the high economic losses that the Company had for this reason and the underutilization of these lands.
The seasonality of the episodes coincides with the fruiting periods of the Canistel that is located in our country in the spring between the months of March - June, González, (1986).
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